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A total of 769 wheat kernels collected from six provinces in China were analyzed for beauvericin (BEA) and four enniatins (ENNs), namely, ENA, ENA1, ENB and ENB1, using a solid phase extraction (SPE) technique with ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The results show that the predominant toxin was BEA, which had a maximum of 387.67 µg/kg and an average of 37.69 µg/kg. With regard to ENNs, the prevalence and average concentrations of ENB and ENB1 were higher than those of ENA and ENA1. The geographical distribution of BEA and ENNs varied. Hubei and Shandong exhibited the highest and lowest positive rates of BEA and ENNs (13.46% and 87.5%, respectively). However, no significant difference was observed among these six provinces. There was a co-occurrence of BEA and ENNs, and 42.26% of samples were simultaneously detected with two or more toxins. Moreover, a significant linear correlation in concentrations was observed between the four ENN analogs (r range: 0.75~0.96, p < 0.05). This survey reveals that the contamination and co-contamination of BEA and ENNs in Chinese wheat kernels were very common.

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Mycotoxins are a class of exogenous metabolites that are major contributors to foodborne diseases and pose a potential threat to human health. However, little attention has been paid to trace mycotoxin co-exposure situations in vivo. To address this, we devised a novel analytical strategy, both highly sensitive and comprehensive, for quantifying 67 mycotoxins in human plasma samples. This method employs isotope dilution mass spectrometry (IDMS) for approximately 40% of the analytes and utilizes internal standard quantification for the rest. The mycotoxins were classified into three categories according to their physicochemical properties, facilitating the optimization of extraction and detection parameters to improve analytical performance. The lowest limits of detection and quantitation were 0.001-0.5 µg/L and 0.002-1 µg/L, respectively, the intra-day precision ranged from 1.8% to 11.9% RSD, and the intra-day trueness ranged from 82.7-116.6% for all mycotoxins except Ecl, DH-LYS, PCA, and EnA (66.4-129.8%), showing good analytical performance of the method for biomonitoring. A total of 40 mycotoxins (including 24 emerging mycotoxins) were detected in 184 plasma samples (89 from infertile males and 95 from healthy males) using the proposed method, emphasizing the widespread exposure of humans to both traditional and emerging mycotoxins. The most frequently detected mycotoxins were ochratoxin A, ochratoxin B, enniatin B, and citrinin. The incidence of exposure to multiple mycotoxins was significantly higher in infertile males than in healthy subjects, particularly levels of ochratoxin A, ochratoxin B, and citrinin, which were significantly increased. It is necessary to carry out more extensive biological monitoring to provide data support for further study of the relationship between mycotoxins and male infertility.

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The total mixed ration (TMR) is currently a widespread method to feed dairy cows. It is a mixture of raw fodder and concentrate feed that can be contaminated by several mycotoxins. The main aim of this paper was to provide a critical review on TMR mycotoxin occurrence and its usefulness to monitor and control them on-farm. Aflatoxins, zearalenone, deoxynivalenol, T-2 toxin and fumonisins (regulated mycotoxins) are the most prevalent mycotoxins evaluated in TMR. Nonetheless, several emerging mycotoxins represent a health risk at the animal level regarding their prevalence and level in TMR. Even when measured at low levels, the co-occurrence of mycotoxins is frequent and synergistic effects on animal health are still underevaluated. Similar to the animal feed industry, on-farm plans monitoring mycotoxin feed contamination can be developed as a herd health management program. The estimated daily intake of mycotoxins should be implemented, but thresholds for each mycotoxin are not currently defined in dairy farms.

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In this work, a complete study of the distribution of emerging mycotoxins in the human body has been carried out. Specifically, the presence of enniatins (A, A1, B, B1) and beauvericin has been monitored in brain, lung, kidney, fat, liver, and heart samples. A unique methodology based on solid-liquid extraction (SLE) followed by dispersive liquid-liquid microextraction (DLLME) was proposed for the six different matrices. Mycotoxin isolation was performed by adding ultrapure water, acetonitrile, and sodium chloride to the tissue sample for SLE, while the DLLME step was performed using chloroform as extraction solvent. Subsequently, the analysis was carried out by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS). The proposed method allowed limits of quantification (LOQs) to be obtained in a range of 0.001-0.150 ng g-1, depending on the tissue and mycotoxin. The precision was investigated intraday and interday, not exceeding of 9.8% of relative standard deviation. In addition, trueness studies achieved 75 to 115% at a mycotoxin concentration of 25 ng g-1 and from 82 to 118% at 5 ng g-1. The application of this methodology to 26 forensic autopsies demonstrated the bioaccumulation of emerging mycotoxins in the human body since all mycotoxins were detected in tissues. Enniatin B (ENNB) showed a high occurrence, being detected in 100% of liver (7 ± 13 ng g-1) and fat samples (0.2 ± 0.8 ng g-1). The lung had a high incidence of all emerging mycotoxins at low concentrations, while ENNB, ENNB1, and ENNA1 were not quantifiable in heart samples. Co-occurrence of mycotoxins was also investigated, and statistical tests were applied to evaluate the distribution of these mycotoxins in the human body.

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Mycotoxins are abiotic hazards whose contamination occurs at the pre- and post-harvest stages of the maize value chain, with animal exposure through contaminated feed leading to their excretion into milk. Currently, only aflatoxin M1 is regulated in milk products. Since feed materials and complete feed present a multi-mycotoxin composition and are the main mycotoxin source into milk, it is important to recognize the occurrence of multiple toxins and their co-occurrence in this highly consumed food product. The aim of this study was to determine the content of regulated and emerging mycotoxins in milk samples, which allowed for evaluating the occurrence and co-occurrence patterns of different mycotoxins known to contaminate feed materials and complete animal feed. Human exposure considering the occurrence patterns obtained was also estimated. Aflatoxins, fumonisins, zearalenone, and emerging mycotoxins were among the mycotoxins found to be present in the 100 samples analyzed. Concentrations ranged from 0.006 to 16.3 µg L-1, with no sample exceeding the AFM1 maximum level. Though several mycotoxins were detected, no exceeding values were observed considering the TDI or PMTDI. It can be concluded that the observed exposure does not pose a health risk to milk consumers, though it is important to recognize vulnerable age groups.

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Although over the last 10 years several studies have focused on the emerging mycotoxins known as enniatins (ENNs), there is still a lack of knowledge regarding their toxicological effects and the development of a correct risk assessment. This is especially true for enniatin B1 (ENN B1), considered the younger sister of the widely studied enniatin B (ENN B). ENN B1 has been found in several food commodities and, as with other mycotoxins, presents antibacterial and antifungal properties. On the other hand, ENN B1 has shown cytotoxic activity, impairment of the cell cycle, the induction of oxidative stress, and changes in mitochondrial membrane permeabilization, as well as negative genotoxic and estrogenic effects. Overall, considering the paucity of information available regarding ENN B1, further studies are necessary to perform a risk assessment. This review summarizes information on the biological characteristics and toxicological effects of ENN B1 as well as the future challenges that this mycotoxin could present.

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Wheat grains are susceptible to contamination with various natural mycotoxins including regulated and emerging mycotoxins. This study surveyed the natural presence of regulated mycotoxins, such as deoxynivalenol (DON) and zearalenone (ZEN), and emerging mycotoxins such as beauvericin (BEA), enniatins (ENNs such as ENA, ENA1, ENB, ENB1) and Alternaria mycotoxins (i.e., alternariol monomethyl ether (AME), alternariol (AOH), tenuazonic acid (TeA), tentoxin (TEN), and altenuene (ALT)) in wheat grains randomly collected from eight provinces across China in 2021. The results revealed that each wheat grain sample was detected with at least one type of mycotoxin. The detection rates of these mycotoxins ranged from 7.1% to 100%, with the average occurrence level ranging from 1.11 to 921.8 µg/kg. DON and TeA were the predominant mycotoxins with respect to both prevalence and concentration. Approximately 99.7% of samples were found to contain more than one toxin, and the co-occurrence of ten toxins (DON + ZEN + ENA + ENA1 + ENB + ENB1 + AME + AOH + TeA + TEN) was the most frequently detected combination. The dietary exposure to different mycotoxins among Chinese consumers aged 4-70 years was as follows: 0.592-0.992 µg/kg b.w./day for DON, 0.007-0.012 µg/kg b.w./day for ZEN, 0.0003-0.007 µg/kg b.w./day for BEA and ENNs, 0.223-0.373 µg/kg b.w./day for TeA, and 0.025-0.041 µg/kg b.w./day for TEN, which were lower than the health-based guidance values for each mycotoxin, with the corresponding hazard quotient (HQ) being far lower than 1, implying a tolerable health risk for Chinese consumers. However, the estimated dietary exposure to AME and AOH was in the range of 0.003-0.007 µg/kg b.w./day, exceeding the Threshold of Toxicological Concern (TTC) value of 0.0025 µg/kg b.w./day, demonstrating potential dietary risks for Chinese consumers. Therefore, developing practical control and management strategies is essential for controlling mycotoxins contamination in the agricultural systems, thereby ensuring public health.

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The recovery of biomolecules from food industry by-products is of major relevance for a circular economy strategy. However, by-products' contamination with mycotoxins represents a drawback for their reliable valorization for food and feed, hampering their application range, especially as food ingredients. Mycotoxin contamination occurs even in dried matrices. There is a need for the implantation of monitoring programs, even for by-products used as animal feed, since very high levels can be reached. This systematic review aims to identify the food by-products that have been studied from 2000 until 2022 (22 years) concerning mycotoxins' contamination, distribution, and prevalence in those by-products. PRISMA ("Preferred Reporting Items for Systematic Reviews and MetaAnalyses") protocol was performed via two databases (PubMed and SCOPUS) to summarize the research findings. After the screening and selection process, the full texts of eligible articles (32 studies) were evaluated, and data from 16 studies were considered. A total of 6 by-products were assessed concerning mycotoxin content; these include distiller dried grain with solubles, brewer's spent grain, brewer's spent yeast, cocoa shell, grape pomace, and sugar beet pulp. Frequent mycotoxins in these by-products are AFB1, OTA, FBs, DON, and ZEA. The high prevalence of contaminated samples, which surpasses the limits established for human consumption, thus limiting their valorization as ingredients in the food industry. Co-contamination is frequent, which can cause synergistic interactions and amplify their toxicity.

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Mycotoxins are compounds produced by several fungi that contaminate agricultural fields and, either directly or by carry-over, final food products. Animal exposure to these compounds through contaminated feed can lead to their excretion into milk, posing threats to public health. Currently, aflatoxin M1 is the sole mycotoxin with a maximum level set in milk by the European Union, as well as the most studied. Nonetheless, animal feed is known to be contaminated by several groups of mycotoxins with relevance from the food safety point of view that can be carried over into milk. To evaluate the multi-mycotoxin occurrence in this highly consumed food product it is crucial to develop precise and robust analytical methodologies towards their determination. In this sense, an analytical method for the simultaneous identification of 23 regulated, non-regulated, and emerging mycotoxins in raw bovine milk using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) was validated. A modified QuEChERS protocol for extraction purposes was used, and further validation was performed by assessing the selectivity and specificity, limits of detection and quantification (LOD and LOQ), linearity, repeatability, reproducibility, and recovery. The performance criteria were compliant with mycotoxin-specific and general European regulations for regulated, non-regulated, and emerging mycotoxins. The LOD and LOQ ranged between 0.001 and 9.88 ng mL-1 and 0.005 and 13.54 ng mL-1, respectively. Recovery values were between 67.5 and 119.8%. The repeatability and reproducibility parameters were below 15 and 25%, respectively. The validated methodology was successfully applied to determine regulated, non-regulated, and emerging mycotoxins in raw bulk milk from Portuguese dairy farms, proving the importance of widening the monitoring scope of mycotoxins in dairy products. Additionality, this method presents itself as a new strategic and integrated biosafety control tool for dairy farms for the analysis of these natural and relevant human risks.

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A proper nutrition is crucial for children's healthy development. Regardless of the usual recommendations to follow a varied diet, some foods can be a source of toxic natural contaminants such as mycotoxins, potent secondary metabolites produced by filamentous fungi. In addition to the most well-known mycotoxins, many of which are subject to tight regulation regarding the maximum levels allowed in different types of food, there is a large group of mycotoxins, the so-called emerging mycotoxins, about which less knowledge has already been acquired, which have gradually been the target of interest from the scientific community due to their prevalence in most foodstuffs, particularly in cereals and cereal-based products. Alternariol and his metabolite alternariol mono-methyl ether, beauvericin, citrinin, culmorin, enniatins, ergot alkaloids, fusaproliferin, kojic acid, moniliformin, sterigmatocystin, tentoxin and tenuazonic acid are the most representative of them. The current review gathered the information of the last ten years that have been published on the levels of emerging mycotoxins in food products dedicated for infants and children. European Union countries are responsible for most of the reported studies, which showed levels that can reach hundreds of mg/kg.

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As the contamination of cereal grains with ergot has been increasing in Western Canada, studies were undertaken to evaluate the impacts of heating (60, 80, 120, or 190 °C) alone or in combination with pelleting on concentrations of ergot alkaloids. Fifteen samples of ergot-contaminated grain from Alberta and Saskatchewan were assayed for R and S epimers of six alkaloids (ergocryptine, ergocristine, ergocornine, ergometrine, ergosine, and ergotamine) using HPLC MS/MS. Five samples with distinct alkaloid profiles were then selected for heating and pelleting studies. Heating resulted in a linear increase (p < 0.05) of total R and total S epimers with increasing temperature, although some individual R epimers were stable (ergometrine, ergosine, ergotamine). Pelleting also increased (p < 0.05) concentrations of total R and total S epimers detected, although ergometrine concentration decreased (p < 0.05) after pelleting. A feeding study arranged in a 2 × 2 factorial structure used 48 backgrounding Angus-cross steers fed four different diets: (1) Control Mash (CM, no added ergot), (2) Control Pellet (CP), (3) Ergot Mash (EM), or (4) Ergot Pellet (EP). Pelleting heated the ergot to 90−100 °C under 4 bars pressure, but the ergot used in the feeding study was not otherwise heated. Alkaloid concentrations of EM and EP varied by up to 1.1 mg/kg depending on the feed matrix assayed. No differences among treatments were noted for growth performance, feed intake, feed conversion, concentrations of serum prolactin and haptoglobin, hair cortisol, or in temperatures of extremities measured by infrared thermography. The only negative impacts of ergot alkaloids were on blood parameters indicative of reduced immune function or chronic inflammation. Pelleting did not heighten the negative clinical outcomes of ergot, although alkaloid concentrations of pelleted feed increased depending on the matrix assayed. It was hypothesized that the heat and pressure associated with pelleting may enhance the recovery of alkaloids from pelleted feed.

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Mycotoxins should be monitored in order to properly evaluate corn silage safety quality. In the present study, corn silage samples (n = 115) were collected in a survey, characterized for concentrations of mycotoxins, and scanned by a NIR spectrometer. Random Forest classification models for NIR calibration were developed by applying different cut-offs to classify samples for concentration (i.e., µg/kg dry matter) or count (i.e., n) of (i) total detectable mycotoxins; (ii) regulated and emerging Fusarium toxins; (iii) emerging Fusarium toxins; (iv) Fumonisins and their metabolites; and (v) Penicillium toxins. An over- and under-sampling re-balancing technique was applied and performed 100 times. The best predictive model for total sum and count (i.e., accuracy mean ± standard deviation) was obtained by applying cut-offs of 10,000 µg/kg DM (i.e., 96.0 ± 2.7%) or 34 (i.e., 97.1 ± 1.8%), respectively. Regulated and emerging Fusarium mycotoxins achieved accuracies slightly less than 90%. For the Penicillium mycotoxin contamination category, an accuracy of 95.1 ± 2.8% was obtained by using a cut-off limit of 350 µg/kg DM as a total sum or 98.6 ± 1.3% for a cut-off limit of five as mycotoxin count. In conclusion, this work was a preliminary study to discriminate corn silage for high or low mycotoxin contamination by using NIR spectroscopy.

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Animal feed (including forage and silage) can be contaminated with mycotoxins. Here, 200 maize silage samples from around China were collected in 2019 and analyzed for regulated mycotoxins, masked mycotoxins (deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, and deoxynivalenol-3-glucoside), and emerging mycotoxins (beauvericin, enniatins, moniliformin, and alternariol). Deoxynivalenol and zearalenone were detected in 99.5% and 79.5% of the samples, respectively. Other regulated mycotoxins were detected in fewer samples. The highest deoxynivalenol and zearalenone concentrations were 3600 and 830 µg/kg, respectively. The most commonly detected masked mycotoxin was 15-acetyldeoxynivalenol, which was detected in 68.5% of the samples and had median and maximum concentrations of 61.3 and 410 µg/kg, respectively. The emerging mycotoxins beauvericin, alternariol, enniatin A, enniatin B1, and moniliformin were detected in 99.5%, 85%, 80.5%, 72.5%, and 44.5%, respectively, of the samples but at low concentrations (medians <25 µg/kg). The samples tended to contain multiple mycotoxins, e.g., the correlation coefficients for the relationships between the concentrations of beauvericin and deoxynivalenol, deoxynivalenol and zearalenone, and zearalenone and beauvericin were 1.0, 0.995, and 0.995, respectively. The results indicated that there needs to be more awareness of the presence of one or more masked and emerging mycotoxins in maize silage in China.

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Emerging mycotoxins, such as enniatin A (ENNA), are becoming a worldwide concern owing to their presence in different types of food and feed. However, comprehensive toxicokinetic data that links intake, exposure and toxicological effects of ENNA has not been elucidated yet. Therefore, the present study investigated the in vitro (rat and human) and in vivo (rat) toxicokinetic properties of ENNA. Towards this, an easily applicable and sensitive bioanalytical method was developed and validated for the estimation of ENNA in rat plasma. ENNA exhibited high plasma protein binding (99%), high hepatic clearance and mainly underwent metabolism via CYP3A4 (74%). The in-house predicted hepatic clearance (54 mL/min/kg) and observed in vivo rat clearance (55 mL/min/kg) were comparable. The predicted in vivo human hepatic clearance was 18 mL/min/kg. ENNA underwent slow absorption (Tmax = 4 h) and rapid elimination following oral administration to rats. The absolute oral bioavailability was 47%. The toxicokinetic findings for ENNA from this study will help in designing and interpreting toxicological studies in rats. Besides, these findings could be used in physiologically based toxicokinetic (PBTK) model development for exposure predictions and risk assessment for ENNA in humans.

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The main objective of this study was to determine the presence of Fusarium metabolites in maize samples collected from different regions of Northern Serbia (Backa, Banat and Srem) during a period of two years (2016-2017). A total of 458 maize samples were analysed by liquid chromatography-tandem mass spectrometry. A total of 40 metabolites were detected, where 94% of the samples contained at least 5 metabolites. Fumonisins (including B1, B2, B3 and B4), moniliformin and bikaverin were the most frequent (80-98%) Fusarium metabolites in both years. Furthermore, in samples from 2016, fumonisin A1 and A2, deoxynivalenol, deoxynivalenol-3-glucoside, zearalenone, culmorin, 15-hydroxyculmorin, fusapyron, fusaproliferin and aurofusarin were detected with frequencies of 58-80%. Levels of certain Fusarium metabolites in 2016 were higher on average due to increased humidity when compared to 2017, which was characterised by warm and dry conditions.

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Exposure to mycotoxins is a worldwide concern as their occurrence is unavoidable and varies among geographical regions. Mycotoxins can affect the performance and quality of livestock production and act as carriers putting human health at risk. Feed can be contaminated by various fungal species, and mycotoxins co-occurrence, and modified and emerging mycotoxins are at the centre of modern mycotoxin research. Preventing mould and mycotoxin contamination is almost impossible; it is necessary for producers to implement a comprehensive mycotoxin management program to moderate these risks along the animal feed supply chain in an HACCP perspective. The objective of this paper is to suggest an innovative integrated system for handling mycotoxins in the feed chain, with an emphasis on novel strategies for mycotoxin control. Specific and selected technologies, such as nanotechnologies, and management protocols are reported as promising and sustainable options for implementing mycotoxins control, prevention, and management. Further research should be concentrated on methods to determine multi-contaminated samples, and emerging and modified mycotoxins.

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The emerging mycotoxin fusaproliferin is produced by Fusarium proliferatum and other related Fusarium species. Several fungi from other taxonomic groups were also reported to produce fusaproliferin or the deacetylated derivative, known as siccanol or terpestacin. Here, we describe the identification and functional characterization of the Fusarium proliferatum genes encoding the fusaproliferin biosynthetic enzymes: a terpenoid synthase, two cytochrome P450s, a FAD-oxidase and an acetyltransferase. With the exception of one gene encoding a CYP450 (FUP2, FPRN_05484), knock-out mutants of the candidate genes could be generated, and the production of fusaproliferin and intermediates was tested by LC-MS/MS. Inactivation of the FUP1 (FPRN_05485) terpenoid synthase gene led to complete loss of fusaproliferin production. Disruption of a putative FAD-oxidase (FUP4, FPRN_05486) did not only affect oxidation of preterpestacin III to terpestacin, but also of new side products (11-oxo-preterpstacin and terpestacin aldehyde). In the knock-out strains lacking the predicted acetyltransferase (FUP5, FPRN_05487) fusaproliferin was no longer formed, but terpestacin was found at elevated levels. A model for the biosynthesis of fusaproliferin and of novel derivatives found in mutants is presented.

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Herbal infusions are amongst the world's most popular and widely enjoyed beverages, due to both large variety and convenience. However, natural contaminants, such as mycotoxins and trace elements can accumulate in aromatic herbs, which may have serious food safety and public health implications. In this study, the presence of mycotoxins, as well as the content of trace elements was evaluated in herbs and herbal infusions commercialized in Brazil. For the determination of fourteen mycotoxins, including the emerging mycotoxins enniatins (EN), beauvericin (BEA), and sterigmatocystin (STE), a liquid-chromatography tandem mass spectrometry (LC-MS/MS) method was validated. Overall, 42 out of 58 herb samples (72%) were contaminated, being BEA the most usual mycotoxin, present in 43% of the samples, followed by STE and HT-2 toxin, present in 37% and 24% of the samples, respectively. In herbal infusions, the occurrence of mycotoxins was 88% lesser than those verified in raw products. Despite these low levels, the hazard quotient (HQ) calculated revealed a potential health concern for HT-2 in infusions. The margin of exposure values for aflatoxins (AF), and ochratoxin A (OTA) from six herbal infusions were below 10,000, indicating also potential health risks. The twenty-one trace elements comprising toxic elements such as arsenic (As), cadmium (Cd), and lead (Pb) were determined in herb raw materials by inductively coupled plasma-mass spectrometry (ICP-MS). The levels of trace elements in herbs were very varied, with aluminum (Al) presenting the highest amount. The levels of legislated elements (As, Cd, Pb) analyzed in herbs were lower than 3.03 µg g-1 (Pb), thus not exceeding the legal limits defined for herbal medicinal by both European and Brazilian pharmacopeia.

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Enniatins are so-called "emerging mycotoxins" that commonly occur in milligrams per kilogram levels in grains and their derived products, as well as in fish, dried fruits, nuts, spices, cocoa, and coffee. The present study investigated the 28-day repeated oral dose toxicity of enniatin complex in CD1(ICR) mice. Enniatin B, enniatin B1, and enniatin A1 at a ratio of 4:4:1 were administered to male and female mice at doses of 0 (vehicle controls), 0.8, 4, and 20 mg/kg body weight/day. In life parameters did not change during the study period, with the exception of slight reductions in food consumption in male mice administered 4 and 20 mg/kg and in female mice administered 20 mg/kg. Body and organ weights did not change, and no alterations in hematology, blood biochemistry, or histopathology parameters were observed at the end of the administration period. Thus, we determined that the no-observed-adverse-effect level of enniatin complex was 20 mg/kg/day for both sexes under the present experimental conditions.

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Paprika is considered a high-quality product being one of the most consumed spices in the world. Contamination with mycotoxins may appear due to inappropriate practices during processing or resulting from invading mould in the final manufactured products. A sample treatment based on dispersive magnetic solid-phase extraction (DMSPE) has been proposed for emerging mycotoxin determination, enniatins (ENNs) and beauvericins (BEAs), in paprika. Different magnetic nanoparticles were tested, and cellulose-ferrite nanocomposite was selected for the extraction and preconcentration of the mycotoxins. Nanocomposite was characterised using field emission scanning electron microscopy and energy dispersive X-ray spectroscopy in terms of morphology and elemental composition. High-resolution mass spectrometry allowed the quantification of the five main emerging mycotoxins and the monitoring of unexpected members of this class of toxic fungal secondary metabolites. The method has been validated, obtaining limits of quantification between 9.5 and 9.9 µg kg-1 and testing its trueness through recovery studies, with satisfactory values of between 89.5 and 97.7%. Relative standard deviations were calculated to evaluate the intra- and inter-day precision and values lower than 8% were obtained in all cases. The analysis of 26 samples, including conventional and organic, demonstrated the presence of ENNB1 at 12.0 ± 0.6 µg kg-1 in one of the samples studied. Other analogues ENNs and BEAs were not detected.