Multi-mycotoxin Determination Research Articles

Method for Determination of Multi-mycotoxins in Milk: QuEChERS Extraction Modified Followed by HPLC-FL Analysis

Method for Determination of Multi-mycotoxins in Milk: QuEChERS Extraction Modified Followed by HPLC-FL Analysis

Determination of multi-mycotoxins in vegetable oil via liquid chromatography-high resolution mass spectrometry assisted by a complementary liquid–liquid extraction

The simultaneous determination of multi-mycotoxins in food commodities are highly desirable due to their potential toxic effects and mass consumption of foods. Herein, liquid chromatography-quadrupole exactive orbitrap mass spectrometry was proposed to analyze multi-mycotoxins in commercial vegetable oils. Specifically, the method featured a successive liquid–liquid extraction process, in which the complementary solvents consisted of acetonitrile and water were optimized. Resultantly, matrix effects were reduced greatly. External calibration approach revealed good quantification property for each analyte. Under optimal conditions, the recovery ranging from 80.8% to 109.7%, relative standard deviation less than 11.7%, and good limit of quantification (0.35 to 45.4 ng/g) were achieved. The high accuracy of proposed method was also validated. The detection of 20 commercial vegetable oils revealed that aflatoxins B1 and B2, zearalenone were observed in 10 real samples. The as-developed method is simple and low-cost, which merits the wide applications for scanning mycotoxins in oil matrices.

Inter-laboratory validation of liquid chromatography-tandem mass spectrometry multi-mycotoxin determination in animal feed - method transfer from the reference laboratory to regional laboratories.

The results are presented of the inter-laboratory validation of a liquid chromatography-tandem mass spectrometry method for the determination of eight mycotoxins (aflatoxin B1, deoxynivalenol, fumonisin B1, fumonisin B2, ochratoxin A, toxin T-2, toxin HT-2 and zearalenone) in animal feeds. This study was an essential part of the method's transfer from the National Reference Laboratory to six regional laboratories in Poland working in the official survey of mycotoxins in feed. The laboratories received a batch of standard solutions, blank samples and quality control materials on which to perform analysis with one procedure and different liquid chromatography-tandem mass spectrometry conditions. The validation results show good precision (reproducibility coefficient of variation 3.7-20.5%) and accuracy of the method (recovery 89-120% and trueness 94-103%) and sufficient skills of the laboratory personnel. The study is an example of the successful transfer of the method among laboratories.

Multi-mycotoxin determination in plant-based meat alternatives and exposure assessment

The aim of this study was to fill in the gap regarding the occurrence of mycotoxins in plant-based meat alternatives. Hence, a multi-mycotoxin method (aflatoxins, ochratoxin A, fumonisins, zearalenone, and mycotoxins from the Alternaria alternata genera) was developed followed by an exposure assessment for the Italian consumers’ exposure to mycotoxins. A total of 13 meat alternatives samples based on soy, pea, chickpea, lupin, and seitan were analysed. With the exception of seitan, all of the remaining samples were contaminated with one mycotoxin or mixtures of up to seven mycotoxins. The level of contamination was as low as 0.2 μg/kg alternariol methyl ether and as high as 66.9 μg/kg fumonisin B1. To analyse the exposure to mycotoxins due to plant-based meat alternatives consumption we used the consumption meat data from the Food and Agriculture Organization for Italian adult consumers and simulated a full replacement of meat with plant-based meat alternatives. Based on our model, consumption of plant-based meat alternatives led to a non-tolerable exposure to alternariol (hazard index (HI) > 1) in pea-based burger and soy + wheat-based steak, while samples contaminated with aflatoxins, respectively ochratoxin A, indicated a health concern related to liver and renal cancer (margin of exposure (MOE) < 10,000). This is the first study that presents the co-occurrence of mycotoxins in multiple plant-based meat alternatives. Moreover, these results indicate that there is a need for policymakers to consider the regulation of mycotoxins in plant-based meat alternatives in order to ensure consumers’ safety.

Risk-benefit assessment of shifting from traditional meat-based diets to alternative dietary patterns.

The consumption of plant-based meat alternatives has recently transitioned from the niche market to the mainstream. However, changing the dietary patterns may lead to new health challenges referred to possible higher exposure to natural toxins found in plant-based food. The present project aimed to train the fellow in the field of chemical risk assessment and provide a comprehensive overview on how a change towards plant-based meat alternatives may represent a driver for emerging risks. Thus, within the EU-FORA programme the fellow engaged in the following activities: (i) perform a systematic review to analyse the distribution of natural toxins in the most common plant-based meat alternatives in Europe, (ii) risk-benefit assessment of shifting from meat-based diets to soy-based meat alternatives by modelling the substitution of meat with soy, and (iii) determine the occurrence of mycotoxins in plant-based meat alternatives from local markets in Parma, Italy. The fellow learnt the chemical risk assessment procedures applied by the researchers from the Department of Food and Drug of Parma University, thus gaining an in-depth expertise in all the steps. A risk-benefit assessment was performed modelling the intake of aflatoxin B1-contaminated soy-based meat analogues. The health impact due to intake of soy and exposure to aflatoxin B1 was estimated. Within the research group, the fellow also worked on developing a multi-mycotoxin determination method for plant-based meat alternatives matrices. The results of the project provide a picture reflecting the occurrence of natural toxins in plant-based meat alternatives and the need of upgraded regulation frameworks that take into account new products and dietary patterns. The EU-FORA fellowship was a great opportunity for the fellow to expand his professional network and increase his expertise in food safety by gaining new skills in chemical risk assessment, risk-benefit assessment and analytical chemistry.

Simultaneous Determination of Alternaria Toxins, Ergot Alkaloid Epimers, and Other Major Mycotoxins in Various Food Matrixes by LC-MS/MS.

Various food commodities are vulnerable to different types of fungal pathogens and could be contaminated with differential classes of mycotoxins as a result. It is ideal to implement a generic method for the simultaneous determination of multi-mycotoxins in different food matrixes or agricultural products. In this study, a simplified sample preparation procedure and a reliable LC-MS/MS analytical method were developed for the comprehensive measurement of 37 regulated and emerging mycotoxins including five Alternaria toxins (ATs) and six major ergot alkaloids (EAs) and their corresponding epimers. Four different food commodities (baby wheat cereal, peanut, tomato puree, and blended flour) were chosen for method validation to demonstrate the applicability of this analytical method across a wide range of food types. Sample extraction was performed using a formic acid-acidified acetonitrile-water (4 + 1, v/v) solution followed by extract dry-down and reconstitution in a water-methanol (1 + 1, v/v) solution for analysis on a biphenyl LC column. Chromatographic analysis was performed using regular acidic LC conditions for baseline separation of ergot alkaloid epimers and completed with a short 11 min cycle time. Accurate quantification was achieved using matrix-matched calibration standards in the range of 0.4 to 400 µg/kg. The recoveries of all mycotoxins (except citrinin) in fortified samples were from 70 to 120%, and the RSD was less than 20%. The established workflow was simple and fast for multi-mycotoxin determination in a wide variety of food commodities with LOQs needed to meet the regulatory levels. The developed method provided the unique benefit of simultaneous analysis of Alternaria toxins (ATs) and ergot alkaloids (EAs) together with other major regulated mycotoxins.

Multi-mycotoxin determination in coffee beans marketed in Tunisia and the associated dietary exposure assessment

Coffee is an agricultural product highly susceptible to mycotoxin contamination. The present study evaluates the natural presence of 17 mycotoxins in different types of coffee sampled in Tunisia (n = 100), by using liquid chromatography-tandem mass spectrometry methods (LC-MS/MS). The results showed that all the analyzed samples were contaminated by at least one mycotoxin. Tentoxin (TENT) was registered in 95 samples with maximum of 300 μg/kg while aflatoxin B1 (AFB1) and ochratoxin A (OTA) in 7 and 18 samples, respectively. Most of OTA positive samples exceeded the OTA maximum regulatory limit (MRL). The highest levels of OTA, aflatoxin G1 (AFG1), and alternariol monomethyl ether (AME) levels were detected in Arabica samples reaching up to 122.6; 186.9 and 388.6 μg/kg, respectively. 82 samples were contaminated by at least two mycotoxins and up to nine different mycotoxins co-occurred simultaneously in two samples. The dietary exposure of the Tunisian population was also assessed, and the probable daily intake (PDI) ranged between 0.27– 1.48 and 2.79–5.28 ng/kg bw/day for OTA and zearalenone (ZEN), respectively. It was also found that, at the worst scenario (Mp), consumers may be reaching up to 2.1% and 8.7% of the tolerable daily intakes (TDIs) of ZEN and OTA, respectively. However, no toxicological risk arises to most of the analyzed mycotoxins from the regular consumption of the evaluated coffee.

Development and Validation of QuEChERS Followed by UHPLC-ToF-MS Method for Determination of Multi-Mycotoxins in Pistachio Nuts.

Pistachios are one of the types of tree nut fruits with the highest mycotoxin contamination, especially of aflatoxins, worldwide. This study developed a Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method that was followed by Ultra-High Performance Liquid Chromatography combined with Time-of-Flight Mass Spectrometry (UHPLC–ToF-MS) for the determination of mycotoxins in pistachios. Different approaches to dispersive solid phase extraction as a clean-up method for high lipid matrices were evaluated. For this, classic sorbents such as C18 (octadecyl-modified silica) and PSA (primary secondary amine), and new classes of sorbents, namely EMR-Lipid (enhanced matrix removal-lipid) and Z-Sep (modified silica gel with zirconium oxide), were used. The QuEChERS method, followed by Z-Sep d-SPE clean-up, provided the best analytical performance for aflatoxins (AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), zearalenone (ZEA), toxin T2 (T2) and toxin HT-2 (HT2) in pistachios. The method was validated in terms of linearity, sensitivity, repeatability, interday precision and recovery; it achieved good results according to criteria imposed by Commission Regulation (EC) no. 401/2006. The method was applied to real samples and the results show that pistachios that are available in Portuguese markets are safe from mycotoxins that are of concern to human health.

Multimycotoxin Determination in Grains: A Comprehensive Study on Method Validation and Assessment of Effectiveness of Controlled Atmosphere Storage in Preventing Mycotoxin Contamination.

Two simple and low-cost QuEChERS approaches were optimized and validated for multimycotoxin determination in grains by UPLC-MS/MS and applied to assess effectiveness of controlled atmosphere (CA) storage in preventing mycotoxin contamination. Common bean, soybean, and maize samples were stored for 6 months. CA treatments were conducted varying O2 and CO2 partial pressures, temperatures, and moisture contents of the chambers. In the validation study for common bean and maize, 8 out of 11 mycotoxins were successfully validated. For soybean, 10 out of 11 mycotoxins were validated. Aflatoxin B1 was detected in all commodities. Statistical tests suggest that storage temperature played a key role in aflatoxin B1 concentrations in common bean and soybean, but had no influence on maize. Maize was also positive for fumonisins B1 and B2. Differences in fumonisin concentrations were not significant among different treatments. Concentrations of aflatoxin B1 in some samples exceeded legislation's maximum levels. Thus, some of the CA treatments applied were effective in preventing mycotoxin contamination in common bean and soybean but were not effective for maize.

Determination of 16 mycotoxins in drug and food homologous products by ultra performance liquid chromatography-tandem mass spectrometry combined with accelerated solvent extraction and QuEChERS

A method was developed for the simultaneous determination of 16 mycotoxins in drug and food homologous products by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with accelerated solvent extraction (ASE) and QuEChERS. The target mycotoxins in drug and food homologous products were extracted by ASE. After concentration, the extracts were purified by QuEChERS. Then, the target compounds were analyzed by UPLC-MS/MS in both positive and negative electrospray ionization and MRM modes. Aflatoxin B1 and fumonisin B1 were quantified by the internal standard method, and the remaining mycotoxins were quantified by the matrix-matched external standard method. The proposed method showed a good linear relationship, with correlation coefficients greater than 0.99. The limits of detection (LODs) and limits of quantification (LOQs) of the 16 mycotoxins ranged from 0.008 μg/kg to 0.3 μg/kg and from 0.03 μg/kg to 1.0 μg/kg, respectively. The blank samples were spiked at three levels, and the recoveries ranged from 70.8% to 118%, with the RSDs being 2.5% to 10.2%. The developed method was successfully applied to mycotoxin analysis in 30 scutellaria, puerarin and sea buckthorn samples bought from local markets. Different levels of mycotoxins were detected in some of the products. The proposed method is simple, rapid and sensitive, and it can be applied to the simultaneous determination of multi-mycotoxins in drug and food homologous products.

Multimycotoxin Determination in Tunisian Farm Animal Feed.

Mycotoxins presence was evaluated in animal feed marketed in Tunisia for the first time ever. A QuEChERS method was performed to analyze the natural copresence of 22 mycotoxins (enniatins, beauvericin, ochratoxin A, aflatoxins, alternariol monomethyl ether, alternariol, tentoxin, zearalenone, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, nivalenol, neosolaniol, diacetoxyscirpenol, T-2 toxin, and HT-2 toxin) in 122 Tunisian marketed feed samples, intended for poultry (n = 43), cattle (n = 35), rabbit (n = 12), sheep (n = 16), and horse (n = 16). Analytes detection and quantification were done using both liquid chromatography and gas chromatography coupled to tandem mass spectrometry. The analytical method showed good linearity (R > 0.996) and sensitivity, the limits of quantification ranged from 0.1ng/g (enniatin A1) to 225ng/g (3-acetyldeoxynivalenol). Eighty-five percent of the analyzed samples were positive. Poultry (n = 43) and rabbit (n = 12) feed samples were the most contaminated. Enniatin B was the most prevalent mycotoxin with values ranged between 0.5ng/g for horse feed and 40ng/g for poultry feed, followed by deoxynivalenol detected from 16ng/g in cattle feed to 250ng/g in poultry feed. None exceeded the limits set by EU recommendations for animal feed. Mycotoxins co-occurrence was observed at most by five different mycotoxins (26%) and up to eight mycotoxins was recorded in 5% of samples. Furthermore, a relatively high copresence rate of different fusariotoxins was registered. Even if no toxicological concern was clearly revealed, the contamination is a real fact and will probably present influence on meat production and on food safety.

Performance Evaluation of LC-MS Methods for Multimycotoxin Determination

Abstract The co-occurrence of regulated mycotoxins in foods and feeds, together with modified (“masked”) and emerging mycotoxins, has been increasingly reported worldwide in recent years. Therefore, sensitive, accurate, and validated methods for the simultaneous determination of these hazardous contaminants in different matrices are highly demanded to fulfil regulatory requirements and to carry out reliable surveillance programs. In these last years, LC-MS methodologies for multimycotoxin screening and/or quantification are being routinely used in control laboratories. However, to date, only one European Standard for multimycotoxin determination is based on LC-MS (EN 16877:2016). The need for standardized LC-MS methods for multimycotoxin determination has been highlighted by regulatory authorities and scientific advisory bodies, including the U.S. Food and Drug Administration and the European Commission. The European Committee for Standardization (CEN) has issued calls for tender for the development of standardized LC-MS methods for mycotoxins in food and animal feeding stuffs. As deliverables, some LC-MS based methods for multimycotoxin determination are currently under approval as European Standards. In addition, the European Commission has recently established specific criteria with which screening methods for mycotoxins, including LC-MS methods, have to comply for use for regulatory purposes. Validation procedures by single-laboratory and collaborative trials have been defined. This paper provides insights and advances on guidelines and tools for performance evaluation of LC-MS methods intended for quantitative determination and for semiquantitative screening of multimycotoxins. In particular, performance criteria set in the European Union and the United States are critically overviewed, and expectations, needs, and future challenges relevant to LC-MS methods for multimycotoxin determination are also discussed.

UHPLC-ToF-MS method for determination of multi-mycotoxins in maize: Development and validation

An Ultra-High Performance Liquid Chromatography combined with Time-of-Flight Mass Spectrometry (UHPLC–ToF-MS) method has been developed for determination of nine mycotoxins, namely aflatoxins (AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), zearalenone (ZEA), toxin T2 (T2) and fumonisins (FB1 and FB2) in maize. The method included a two-step extraction with acetonitrile 80% (v/v). After optimization, the analytical method was validated. The different concentrations tested take in account the Maximum Levels (ML) for maize (Commission Regulation EC no. 1881/2006) and good results for repeatability (%RSDr ≤ 15.4%), reproducibility (%RSDR ≤ 15.9%) and recovery (77.8–110.4%, except for AFG2 at 2 μg/kg which presented a recovery of 73.4%) were achieved. These met the performance criteria imposed by Commission Regulation (EC) no. 401/2006. The method was applied to twenty-two samples from Portuguese producers of maize. Fumonisins were the most frequently detected mycotoxins, but the levels do not exceed those imposed by European legislation.

A Dilute-and-Shoot Based UPLC-MS/MS Method for Multimycotoxins Analysis in Astragali radix.

Background: Astragali radix is prone to be contaminated by various mycotoxins, leading to unpredictable threats on the quality and safety of Astragali radix and the health of the consumers. Therefore, the determination of multimycotoxins is imperative. Objective: To develop an efficient, sensitive, fast, and multianalyte method for detecting multimycotoxins in Astragali radix. Methods: A selective dilute-and-shoot pretreatment procedure based ultra-performance LC-tandem MS (UPLC-MS/MS) method was developed for sensitive determination of multimycotoxins in Astragali radix, including aflatoxin (AF) B1, B₂, G1, G₂, and ochratoxin A. The five mycotoxins were extracted by the dilute-and-shoot pretreatment procedure followed by UPLC-MS/MS detection. Types of sample extraction solvent, mobile-phase compositions and MS/MS parameters, and dilute-and-shoot conditions were optimized. Results: The optimized chromatographic and mass spectrometric conditions allowed the separation and detection of the five mycotoxins within 5 min. The validated UPLC-MS/MS method exhibited good sensitivity with LOD and LOQ lower than 0.2 and 0.5 μg/kg, respectively. RSD values for method precision were lower than 9%. Recoveries obtained were between 90.87 and 108.44% for all the analytes with RSDs of 3.96-8.03%. The developed method was applied for the detection of the 5 mycotoxins in 18 batches of Astragali radix with good determination performance and no matrix interferences. Two samples collected from Shanxi province and Neimenggu Autonomous Region in China were positive for AFB1 at 3.24 and 2.69 μg/kg, respectively. Conclusions: The dilute-and-shoot procedure allowed for the extraction of the mycotoxins with advantages of simple pretreatment, small extraction time, high selectivity and accuracy, as well as being cost effective and easy to operate without any clean-up steps. Highlights: This is the first report on dilute-and-shoot approach for mycotoxins extraction and detection in Chinese medicinal material matrixes.

Performance Evaluation of LC-MS Methods for Multimycotoxin Determination.

The co-occurrence of regulated mycotoxins in foods and feeds, together with modified ("masked") and emerging mycotoxins, has been increasingly reported worldwide in recent years. Therefore, sensitive, accurate, and validated methods for the simultaneous determination of these hazardous contaminants in different matrices are highly demanded to fulfil regulatory requirements and to carry out reliable surveillance programs. In these last years, LC-MS methodologies for multimycotoxin screening and/or quantification are being routinely used in control laboratories. However, to date, only one European Standard for multimycotoxin determination is based on LC-MS (EN 16877:2016). The need for standardized LC-MS methods for multimycotoxin determination has been highlighted by regulatory authorities and scientific advisory bodies, including the U.S. Food and Drug Administration and the European Commission. The European Committee for Standardization (CEN) has issued calls for tender for the development of standardized LC-MS methods for mycotoxins in food and animal feeding stuffs. As deliverables, some LC-MS based methods for multimycotoxin determination are currently under approval as European Standards. In addition, the European Commission has recently established specific criteria with which screening methods for mycotoxins, including LC-MS methods, have to comply for use for regulatory purposes. Validation procedures by single-laboratory and collaborative trials have been defined. This paper provides insights and advances on guidelines and tools for performance evaluation of LC-MS methods intended for quantitative determination and for semiquantitative screening of multimycotoxins. In particular, performance criteria set in the European Union and the United States are critically overviewed, and expectations, needs, and future challenges relevant to LC-MS methods for multimycotoxin determination are also discussed.

The contribution of lot-to-lot variation to the measurement uncertainty of an LC-MS-based multi-mycotoxin assay

Multi-mycotoxin determination by LC-MS is commonly based on external solvent-based or matrix-matched calibration and, if necessary, the correction for the method bias. In everyday practice, the method bias (expressed as apparent recovery RA), which may be caused by losses during the recovery process and/or signal/suppression enhancement, is evaluated by replicate analysis of a single spiked lot of a matrix. However, RA may vary for different lots of the same matrix, i.e., lot-to-lot variation, which can result in a higher relative expanded measurement uncertainty (Ur). We applied a straightforward procedure for the calculation of Ur from the within-laboratory reproducibility, which is also called intermediate precision, and the uncertainty of RA (ur,RA). To estimate the contribution of the lot-to-lot variation to Ur, the measurement results of one replicate of seven different lots of figs and maize and seven replicates of a single lot of these matrices, respectively, were used to calculate Ur. The lot-to-lot variation was contributing to ur,RA and thus to Ur for the majority of the 66 evaluated analytes in both figs and maize. The major contributions of the lot-to-lot variation to ur,RA were differences in analyte recovery in figs and relative matrix effects in maize. Ur was estimated from long-term participation in proficiency test schemes with 58%. Provided proper validation, a fit-for-purpose Ur of 50% was proposed for measurement results obtained by an LC-MS-based multi-mycotoxin assay, independent of the concentration of the analytes.

Multi-mycotoxin determination in rice, maize and peanut products most consumed in Côte d’Ivoire by UHPLC-MS/MS

The aim of this study was to determine the multi-mycotoxin occurrence in cereal and oilseed products most consumed in Côte d’Ivoire. A total of 238 samples of rice (88 produced locally or imported), maize (79, cracked or flour) and peanut paste (71) were collected in the main markets of Abidjan, Bouaké and Korhogo. An UHPLC-MS/MS method allowed the analysis of 77 mycotoxins. All the peanut paste samples were contaminated by aflatoxin B1 (AFB1) with 99% exceeding the EU limits of 2 μg kg−1 for AFB1 and 4 μg kg−1 for total aflatoxins (AFT: B1+B2+G1+G2), and concentrations reaching up to 4535 μg kg−1 (AFB1) and 8094 μg kg−1 (AFT). Maize (96%) and rice (57%) samples were also contaminated by AFB1 with 58% and 24% respectively above the EU limits and maximum levels of 80 μg kg−1 for maize and 14 μg kg−1 for rice. Only 6% of the cereal samples (3 rice and one maize samples) had ochratoxin A content above the EU limit (3 μg kg−1). Fumonisins and zearalenone were detected, respectively, in 91% and 8% of the maize samples, and in 18% and 5% of the rice samples but at levels below EU limits. Out of the 238 samples, 91% were contaminated with more than one mycotoxin including EU regulated mycotoxins and/or other mycotoxins mainly beauvericin (79% of the samples), equisetin (71%), aflatoxin M1 (45%), cyclopiazonic acid (32%), fumonisin B3 (29%), sterigmatocystin (24%), citrinin (18%), ochratoxin B (16%) and fusaric acid (15%). The peanut paste samples represented the highest risk to consumer health followed by maize and rice samples.

Rapid Detection and Identification of Mycotoxigenic Fungi and Mycotoxins in Stored Wheat Grain.

This study aimed to assess the occurrence of toxigenic fungi and mycotoxin contamination in stored wheat grains by using advanced molecular and analytical techniques. A multiplex polymerase chain reaction (PCR) strategy was established for rapid identification of mycotoxigenic fungi, and an improved analytical method was developed for simultaneous multi-mycotoxin determination in wheat grains by liquid chromatography-tandem mass spectrometry (LC/MS/MS) without the need for any clean-up. The optimized multiplex PCR method was highly specific in detecting fungal species containing species-specific and mycotoxin metabolic pathway genes. The method was applied for evaluation of 34 wheat grain samples collected from storage warehouses for the presence of mycotoxin-producing fungi, and a few samples were found positive for Fusarium and Aspergillus species. Further chemical analysis revealed that 17 samples contained mycotoxins above the level of detection, but only six samples were found to be contaminated over the EU regulatory limits with at least one mycotoxin. Aflatoxin B1, fumonisins, and deoxynivalenol were the most common toxins found in these samples. The results showed a strong correlation between the presence of mycotoxin biosynthesis genes as analyzed by multiplex PCR and mycotoxin detection by LC/MS/MS. The present findings indicate that a combined approach might provide rapid, accurate, and sensitive detection of mycotoxigenic species and mycotoxins in wheat grains.

Development of a LC-MS/MS Method for the Multi-Mycotoxin Determination in Composite Cereal-Based Samples.

The analytical scenario for determining contaminants in the food and feed sector is constantly prompted by the progress and improvement of knowledge and expertise of researchers and by the technical innovation of the instrumentation available. Mycotoxins are agricultural contaminants of fungal origin occurring at all latitudes worldwide and being characterized by acute and chronic effects on human health and animal wellness, depending on the species sensitivity. The major mycotoxins of food concern are aflatoxin B1 and ochratoxin A, the first for its toxicity, and the second for its recurrent occurrence. However, the European legislation sets maximum limits for mycotoxins, such as aflatoxin B1, ochratoxin A, deoxynivalenol, fumonisins, and zearalenone, and indicative limits for T-2 and HT-2 toxins. Due to the actual probability that co-occurring mycotoxins are present in a food or feed product, nowadays, the availability of reliable, sensitive, and versatile multi-mycotoxin methods is assuming a relevant importance. Due to the wide range of matrices susceptible to mycotoxin contamination and the possible co-occurrence, a multi-mycotoxin and multi-matrix method was validated in liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the purpose to overcome specific matrix effects and analyze complex cereal-based samples within the Italian Total Diet Study project.

Simultaneous Determination of Multi-Mycotoxins in Cereal Grains Collected from South Korea by LC/MS/MS

An improved analytical method compared with conventional ones was developed for simultaneous determination of 13 mycotoxins (deoxynivalenol, nivalenol, 3-acetylnivalenol, aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, fumonisin B1, fumonisin B2, T-2, HT-2, zearalenone, and ochratoxin A) in cereal grains by liquid chromatography-tandem mass spectrometry (LC/MS/MS) after a single immunoaffinity column clean-up. The method showed a good linearity, sensitivity, specificity, and accuracy in mycotoxin determination by LC/MS/MS. The levels of 13 mycotoxins in 5 types of commercial grains (brown rice, maize, millet, sorghum, and mixed cereal) from South Korea were determined in a total of 507 cereal grains. Mycotoxins produced from Fusarium sp. (fumonisins, deoxynivalenol, nivalenol, and zearalenone) were more frequently (more than 5%) and concurrently detected in all cereal grains along with higher mean levels (4.3–161.0 ng/g) in positive samples than other toxins such as aflatoxins and ochratoxin A (less than 9% and below 5.2 ng/g in positive samples) from other fungal species.