Study of co-occurrence of mycotoxins in cocoa beans in Brazil by liquid chromatography-tandem mass spectrometry (LC-MS/MS).

In this study, 135 samples of cocoa beans collected in the Amazon and Atlantic Forest regions of Brazil were analysed to evaluate the possible co-occurrence of 34 mycotoxins. The results indicate that 42% of the cocoa samples exhibited quantifiable levels for 11 mycotoxins: aflatoxins (AFs) B1, B2 and G1; ochratoxin A; citrinin; cyclopiazonic acid; tenuazonic acid; paxilline; sterigmatocystin; zearalenone and fumonisin B2. Of the samples, 18% exhibited the co-occurrence of up to six mycotoxins. No toxins belonging to the groups of trichothecenes or ergot alkaloids were detected. Contingency analysis of the incidence of mycotoxins did not show significant differences between the two regions evaluated. Seven samples were contaminated with AFs, while only one contained ochratoxin A above 10 µg kg-1. The accuracy of the method was evaluated by proficiency testing for ochratoxin A, where satisfactory Z-scores were obtained.

Co-Occurrence of Mycotoxins in Feed for Cattle, Pigs, Poultry, and Sheep in Navarra, a Region of Northern Spain.

Mycotoxins, toxic compounds produced by fungi on raw materials, such as cereals, represent a serious health hazard. Animals are exposed to them mainly through the ingestion of contaminated feed. This study presents data about the presence and co-occurrence of nine mycotoxins: aflatoxins B1, B2, G1, and G2, ochratoxins A and B, zearalenone (ZEA), deoxynivalenol (DON), and sterigmatocystin (STER), in 400 samples of compound feed for cattle, pigs, poultry, and sheep (100 samples each) collected in Spain (2019-2020). Aflatoxins, ochratoxins, and ZEA were quantified using a previously validated HPLC method using fluorescence detection; whereas DON and STER were quantified using ELISA. Moreover, the obtained results were compared with those obtained in this country and published in the last 5 years. The mycotoxin presence in Spanish feed, especially for ZEA and DON, has been demonstrated. The maximum individual levels found were: AFB1: 6.9 µg/kg in a sample of feed for poultry; OTA: 65.5 µg/kg in a sample of feed for pigs, DON: 887 µg/kg in a sample of feed for sheep, and ZEA: 816 µg/kg in a sample of feed for pigs. Nevertheless, regulated mycotoxins appear, in general, at levels below those regulated by the EU; in fact, the percentage of samples containing concentrations above these limits was very low (from 0% for DON to 2.5% for ZEA). The co-occurrence of mycotoxins has also been demonstrated: 63.5% of the analyzed samples presented detectable levels of two to five mycotoxins. Due to the fact that the distribution of mycotoxins in raw materials can change greatly from year to year with climate conditions or market globalization, regular mycotoxin monitorization in feed is needed to prevent the integration of contaminated materials in the food chain.

[Establishment of non-targeted screening database and confirmation method for 18 mycotoxins in grains using ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry].

A mass spectral library of 18 mycotoxins was developed based on ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF/MS), which was used to establish a non-targeted screening method for mycotoxins in rice and wheat matrices. Eighteen mycotoxin standards were separated on an HSS T3 column, and data were collected for both positive and negative ionization under the MSE mode of the UPLC-Q-TOF/MS. Details including formulas, retention times, theoretical exact masses, measured exact masses of the adduct and fragment ions, and ion abundance ratios were recorded to establish the mass spectral library of the 18 mycotoxins in UNIFI Software. Analyte detection was based on a retention time deviation of 0.3 min, and the exact mass deviation of the adduct ions and fragment ions was set to 5×10-6. The screening detection limit (SDL) was used as the main threshold for verifying the screening method. In the validation process, 18 mycotoxins were classified into two types: with maximum levels (MLs) and without MLs. The results showed that the mycotoxins with MLs could be accurately screened at their limited level, and the mycotoxins without MLs had a range of SDL concentration from 2 to 800 µg/kg. The matrix effect results showed that 14 mycotoxins in rice and 11 in wheat had moderate matrix effects. Finally, 25 batches of rice and wheat were purified using QuEChERS and HLB columns after acetonitrile extraction and screening were performed by employing the established method. The results revealed that aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), fumonisins B1 (FB1), and sterigmatocystin (ST) were detected in one batch of rice, FB1 and ST were detected in another batch of rice, FB1 and ochratoxin A (OTA) were detected in two batches of wheat, and no other mycotoxins were detected. This method is characterized by high throughput, simplicity, rapidity, accuracy, and can be applied to accurately screen mycotoxins with concentrations higher than the SDLs and qualitatively screen various mycotoxins in rice and wheat without standards.

Centrifugation-Assisted Solid-Phase Extraction Coupled with UPLC-MS/MS for the Determination of Mycotoxins in ARECAE Semen and Its Processed Products.

Mycotoxins can occur naturally in a variety of agriculture products, including cereals, feeds, and Chinese herbal medicines (TCMs), via pre- and post-harvest contamination and are regulated worldwide. However, risk mitigation by monitoring for multiple mycotoxins remains a challenge using existing methods due to their complex matrices. A multi-toxin method for 22 mycotoxins (aflatoxin B1, B2, G1, G2, M1, M2; ochratoxin A, B, C; Fumonisin B1, B2, B3; 15-acetyldeoxynivalenol, 3-acetyldeoxynivalenol, diace-toxyscirpenol, HT-2, T-2, deepoxy-deoxynivalenol, deoxynivalenol, neosolaniol, zearalenone, and sterigmatocystin) using centrifugation-assisted solid-phase extraction (SPE) clean-up prior to ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis for Arecae Semen and its processed products was developed and validated. Several experimental parameters affecting the extraction and clean-up efficiency were systematically optimized. The results indicated good linearity in the range of 0.1-1000 µg/kg (r2 > 0.99), low limits of detection (ranging from 0.04 µg/kg to 1.5 µg/kg), acceptable precisions, and satisfactory recoveries for the selected mycotoxins. The validated method was then applied to investigate mycotoxin contamination levels in Areca catechu and its processed products. The mycotoxins frequently contaminating Areca catechu were aflatoxins (AFs), and the average contamination level and number of co-occurring mycotoxins in the Arecae Semen slices (Binlangpian) were higher than those in commercially whole Arecae Semen and Arecae Semen Tostum (Jiaobinlang). Sterigmatocystin was detected in 5 out of 30 Arecae Semen slices. None of the investigated mycotoxins were detected in Arecae pericarpium (Dafupi). The results demonstrated that centrifugation-assisted SPE coupled with UHPLC-MS/MS can be a useful tool for the analysis of multiple mycotoxins in Areca catechu and its processed products.

Enzyme immunoassays for the detection of mycotoxins in plant-based milk alternatives: pitfalls and limitations.

Plant-based milk alternatives (PBMAs) are a potential source of mycotoxin uptake. To ensure food safety, simple and rapid testing methods of PBMAs for mycotoxins are therefore required. This study investigated the applicability of enzyme immunoassay (EIA) methods for direct testing of PBMAs without sample extraction. Mycotoxin analyses included aflatoxin B1 (AFB1), sterigmatocystin (STC), ochratoxin A (OTA), deoxynivalenol (DON), and T-2/HT-2-toxin (T-2/HT-2). It was found that the PBMA matrix negatively affected the EIA to varying degrees, thus affecting the reliability of the results. A dilution of PBMAs of at least 1:8 was necessary to overcome matrix interference. This resulted in calculated detection limits of 0.4 µg/L (AFB1), 2 µg/L (STC), 0.08 µg/L (OTA), 16 µg/L (DON), and 0.4 µg/L (T-2/HT-2). After analysis of 54 PBMA products from German retail stores, positive results in at least one test system were obtained for 23 samples. However, most positive results were near the calculated detection limit. Control analyses of selected samples by LC-MS/MS for AFB1, STC, and OTA qualitatively confirmed the presence of trace amounts of STC in some samples, but quantitative agreement was poor. It was concluded that the high diversity of ingredients used in PBMAs led to a highly variable degree of sample matrix interference even in a 1:8 dilution. Since the use of higher dilutions conflicts with the need to achieve low detection limits, the application of EIA for routine mycotoxin analysis in PBMA for mycotoxins requires further study on the development of a feasible sample preparation method.

Ochratoxin A and Sterigmatocystin in Long-Ripened Grana Cheese: Occurrence, Wheel Rind Contamination and Effectiveness of Cleaning Techniques on Grated Products.

A survey on the occurrence of ochratoxin A (OTA) and sterigmatocystin (STC) in grated cheese products obtained from hard grana-type cheeses was carried out, where 107 grated products were collected in retail outlets and analysed. OTA and STC were found in 48.6% and 94.4% of the samples, in a range from

Profiles of Sterigmatocystin and Its Metabolites during Traditional Chinese Rice Wine Processing.

Mycotoxin pollution is widespread in cereal, which greatly threatens food security and human health. In this study, the migration and transformation of sterigmatocystin (STG) mycotoxin during the contaminated rice wine processing was systematically assessed. QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) coupled with ultrahigh-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) method was firstly established for STG analysis in rice wine. It was found that high levels of rice leaven caused a significant reduction in STG in the fermented rice and wine, which was mainly due to the adsorption of yeast cells and Rhizopus biological degradation. However, compared with rice, the levels of STG in separated fermented wine was significantly decreased by 88.6%, possibly attributed to its high log Kow (3.81) and low water solubility (1.44 mg/L). The metabolites of STG (i.e., monohydroxy STG) were identified in rice wine fermentation for the first time. Moreover, STG disturbed the metabolic profile rice wine composition mainly by glycine, serine and threonine metabolism, alanine, aspartate and glutamate metabolism, purine metabolism pathway, particularly with regard to eight amino acids and sixteen lipids. This study elucidated the STG migration and transformation mechanism during the rice wine processing. The finding provided new analytical method for mycotoxin exposure and pollutant in food production, which may support agricultural production and food security.

Occurrence and risk assessment of sterigmatocystin in agricultural products and processed foods in Korea.

Sterigmatocystin (STC), a carcinogenic mycotoxin, is known to be produced during the biosynthetic pathway of aflatoxin B1. STC in various foods was determined by LC-MS/MS and its risks were assessed. The analytical method was validated in different food categories, and the performance was acceptable based on the criteria of AOAC. A total 1,135 samples (613 agricultural products and 522 processed foods) were analysed, and STC was detected in 46 samples, indicating a detection rate of 4.1%. STC was found in the range of 0.08-10.07 ng/g, and the detection rates of STC were 3.9% in agricultural products and 4.2% in processed foods. The exposure to STC by average consumption of foods was estimated to 0.09 ng/kg b.w./day. The margin of exposure (MOE) approach was applied to assess the risk of STC, and MOE for the whole population was over 1 × 106. Exposure to STC from the consumption of foods distributed in Korea is unlikely to cause human health problems.

Aflatoxin B1 and sterigmatocystin: method development and occurrence in tea.

Tea is one of the most popular beverage in the world and may be contaminated by fungi and mycotoxins during processing. To analyse aflatoxin B1 (AFB1) and sterigmatocystin (STC) in three types of tea, a simple, fast, sensitive and reliable method of these two myxotoxins was developed. Recoveries obtained ranged from 95.9% to 118.0% and the RSDs were between 0.3% and 11.2%. The range of LODs was 0.2-0.45 µg/kg for AFB1 and 0.04-0.12 µg/kg for STC. The range of LOQs was 0.67-1.73 µg/kg for AFB1 and 0.13-0.40 µg/kg for STC. The optimised procedure was applied to analyse 126 tea samples randomly collected from different markets in China. AFB1 was not detected, but STC was determined in 17 samples with concentrations ranging from 0.13 to 4.48 µg/kg. The detection rate of STC was 5%, 8.9% and 33.3% in black tea, green tea and Oolong tea, respectively.

Biomonitoring of Mycotoxins in Plasma of Patients with Alzheimer's and Parkinson's Disease.

Exposure to environmental contaminants might play an important role in neurodegenerative disease pathogenesis, such as Parkinson´s disease (PD) and Alzheimer´s disease (AD). For the first time in Spain, the plasmatic levels of 19 mycotoxins from patients diagnosed with a neurodegenerative disease (44 PD and 24 AD) and from their healthy companions (25) from La Rioja region were analyzed. The studied mycotoxins were aflatoxins B1, B2, G1, G2 and M1, T-2 and HT-2, ochratoxins A (OTA) and B (OTB), zearalenone, sterigmatocystin (STER), nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deepoxy-deoxynivalenol, neosolaniol, diacetoxyscirpenol and fusarenon-X. Samples were analyzed by LC-MS/MS before and after treatment with ß-glucuronidase/arylsulfatase in order to detect potential metabolites. Only OTA, OTB and STER were detected in the samples. OTA was present before (77% of the samples) and after (89%) the enzymatic treatment, while OTB was only detectable before (13%). Statistically significant differences in OTA between healthy companions and patients were observed but the observed differences might seem more related to gender (OTA levels higher in men, p-value = 0.0014) than the disease itself. STER appeared only after enzymatic treatment (88%). Statistical analysis on STER, showed distributions always different between healthy controls and patients (patients' group > controls, p-value < 0.0001). Surprisingly, STER levels weakly correlated positively with age in women (rho = 0.3384), while OTA correlation showed a decrease of levels with age especially in the men with PD (rho = -0.4643).

A New Protocol for the Detection of Sterigmatocystin-producing Aspergillus Section Versicolores Using a High Discrimination Polymerase.

Aspergillus section Versicolores species, except Aspergillus sydowii, produce a carcinogenic mycotoxin sterigmatocystin (STC). Since these fungi are found in varied environmental milieu including indoor dust and food products, our aim was to develop a sensitive and convenient assay to detect STC producing fungal strains. We made use of a high discrimination DNA polymerase (HiDi DNA polymerase), for single nucleotide polymorphism (SNP)-based PCR amplification. Using specific primer pairs based on the SNPs between A. sydowii and other strains of Aspergillus section Versicolores, we succeeded in amplifying the genomic DNA all target strains except A. sydowii. These results confirm that the SNP-based PCR amplification technique, using a high discrimination DNA polymerase, was a reliable and robust screening method for target fungal strains.

Selective recognition and enrichment of sterigmatocystin in wheat by thermo-responsive imprinted polymer based on magnetic halloysite nanotubes.

Two thermo-responsive molecularly imprinted polymers (MHNTs@MIP and MCNTs@MIP) for the selective extraction of sterigmatocystin have been prepared on the surface of the magnetic halloysite nanotubes (MHNTs) and magnetic carbon nanotubes (MCNTs), respectively. 1, 8-dihydroxyanthraquinone, n-isopropyl acrylamide, methacrylic acid, ethylene dimethacrylate and dimethyl sulfoxide were used as the dummy template, thermo-sensitive functional monomer, co-monomer, cross-linker and porogen, respectively. The magnetic properties, adsorption properties as well as the temperature responsive behaviors of MHNTs@MIP and MCNTs@MIP were systematically studied and compared for the first time. Enough saturation magnetizations of MHNTs@MIP (9.42 emu/g) and MCNTs@MIP (10.54 emu/g) were obtained. MHNTs@MIP and MCNTs@MIP also showed controllable adsorption and release behaviors to sterigmatocystin in response to the temperature change (35 °C and 20 °C). Compared with MCNTs@MIP, MHNTs@MIP had higher adsorption affinity (KL = 0.120 L/mg), higher adsorption kinetic (K2 = 0.0100 g/(mg•min)) and higher imprinting factor (5.22) to sterigmatocystin. These results indicated that MHNTs@MIP was favorable adsorbent for the selective separation of sterigmatocystin. Furthermore, the elution conditions of MHNTs@MIP were optimized by response surface methodology. Under the optimal conditions, MHNTs@MIP coupled with high performance liquid chromatography were successfully applied to the selective recognition, purification, enrichment and detection of sterigmatocystin in wheat samples. The recoveries were calculated from 88.62% to 102.9% with RSDs less than 3.5 % and limit of detection of 1.1 µg/kg. This work provided a suitable carrier for the preparation of imprinted polymers and a practical approach for highly selective recognition and determination of analytes in real samples.

Development and validation of an analytical method for the analysis of Sterigmatocystin in roasted coffee beans and black pepper using liquid chromatography-tandem mass spectrometry.

Sterigmatocystin (STC) is a toxic and potentially carcinogenic fungal toxin found in a variety of food commodities. This study describes the development of an analytical method to determine STC in roasted coffee beans and black pepper using ultra performance liquid chromatography (UPLC) coupled with triple quadrupole tandem mass spectrometry (MS/MS). 13C18-STC was used as internal standard. STC was extracted with a mixture of acetonitrile/water, diluted with a buffer, followed by purification with a solid-phase extraction and an immunoaffinity column prior to the UPLC-MS/MS analysis. Two multiple reaction monitoring (MRM) transitions were employed, one for quantification and one for confirmation of STC. The UPLC-MS/MS analytical method was validated with respect to selectivity, linearity, sensitivity, accuracy, precision, recovery, and stability. Calibration curves were linear over a concentration range 25-2,500 pg mL-1 with correlation coefficients (r) > 0.998. The method limit of quantification for STC in roasted coffee beans and black pepper was 0.10 µg kg-1. The accuracy and precision of the analytical method were acceptable within 15% at all quality control levels. This method was suitable to determine STC levels because of its selectivity, precision, and accuracy. The method was successfully applied to roasted coffee beans and black pepper samples.

Determination of sterigmatocystin in foods in Japan: method validation and occurrence data.

A survey of the contamination of foods by sterigmatocystin (STC) was performed by an analytical method based on LC-MS/MS. STC was extracted from samples with acetonitrile/water (85/15, v/v) and then purified with immunoaffinity columns. The method was validated by a small-scale inter-laboratory study using spiked wheat samples. Mean recoveries of STC were 100.3% and 92.5% from two samples spiked at 0.5 and 5.0 µg/kg, respectively. A total of 583 samples were analysed between 2016 and 2018, and STC was detected in 19.9% of all samples at >0.05 µg/kg (limit of quantification). The foods that were contaminated by STC were wheat flour, Job's tears products, rye flour, rice, buckwheat flour, white sorghum, barley products, azuki bean and corn flour. STC was not found in beer or wine. The occurrence of STC in domestic wheat flour (44.4%), Job's tears products (41.7%) and rye flour (29.9%) accounted for the three highest values. The highest mean concentrations were obtained for Job's tears products (0.3 µg/kg) and rye flour (0.3 µg/kg). The maximum contamination level was present in a sample of rye flour (7.1 µg/kg). Although the contamination levels were low, these results indicate that STC frequently contaminates Japanese retail foods. A continuous survey is required to assess exposure to STC in Japan.

Development of a polyclonal antibody-based indirect competitive ELISA for determination of sterigmatocystin in wheat and corn flours.

Sterigmatocystin (STC) is a toxic secondary metabolite produced by more than 50 fungal species, including Aspergillus flavus, A. parasiticus, A. nidulans, and A. versicolor. The Joint FAO/WHO Expert Committee on Food Additives concluded that sterigmatocystin is genotoxic and carcinogenic with the critical effect determined to be carcinogenicity. The present study describes a simple method to prepare hapten and immunogens in order to generate polyclonal antibodies against this metabolite. We developed a sensitive and specific polyclonal antibody-based competitive indirect enzyme-linked immunosorbent assay (ciELISA) for monitoring STC in wheat and corn flours without the need for derivatisation of STC or clean-up of samples by immunoaffinity chromatography for quantification. The half inhibitory concentration (IC50) of the established method was 4.52 ± 0.81 ng mL-1, with the limit of detection (IC10) being 0.19 ± 0.04 ng mL-1 in wheat and corn flour matrices with the coefficient of variation of less than 22%.The assay was very specific to STC and showed no cross-reactivity with its analogue structures. The ELISA allowed for up to 5% methanol without significant influence on the IC50 value. Validation of the assay was performed by spiking STC into a blank flour matrix and the recoveries were in the range of 75.3 % to 104.5 % with a coefficient of variation less than 15%. A small retail survey was conducted by purchasing wheat (n = 8) and corn flours (n = 2) from local grocery stores. All of these retail samples were negative for STC using the developed ELISA method and were confirmed by LC-MS/MS. We demonstrated a rapid, simple, and reliable method for screening STC in wheat and corn flours.

Development of a third-generation biosensor to determine sterigmatocystin mycotoxin: An early warning system to detect aflatoxin B1.

A third-generation enzymatic biosensor was developed to quantify sterigmatocystin (STEH). It was based on a glassy carbon electrode modified with a composite of the soybean peroxidase enzyme (SPE) and chemically reduced graphene oxide. The optimal conditions to construct the biosensor were obtained through an experimental design based on the response surfaces methodology. The experiments were performed in 0.1 mol L-1 phosphate buffer solution, pH 5. Amperometric measurements were carried out at - 0.09 V vs Ag/AgCl (3 mol L-1 NaCl). The biosensor showed a lineal response in the concentration range from 6.9 × 10-9 to 5.0 × 10-7 mol L-1. The limit of detection was 2.3 × 10-9 mol L-1 for a signal: noise ratio of 3: 1. Values of the apparent Michaellis-Menten constant, KMapp, obtained by using both Lineweaver-Burk and Eadi-Hofstee methods were (1.5 ± 0.2) × 10-6 and (1.2 ± 0.2) × 10-6 mol L-1, respectively. STEH was analyzed in corn samples spiked with STEH, with an average recovery of 96.5%. The biosensor was also used to determine STEH in corn samples inoculated with the Aspergillus flavus fungus, which is an aflatoxins producer. Considering that STEH is a precursor of aflatoxin B1 (AFB1) in its biological transformation, its decrease over time was related to the production of AFB1. The STEH concentration determined using the biosensor was in very good agreement with that determined by HPLC.

Optimization of Storage Conditions of the Medicinal Herb Ilex asprella against the Sterigmatocystin Producer Aspergillus versicolor Using Response Surface Methodology.

The root of Ilex asprella is a commonly used herb in Southern China, and also constitutes the main raw material of Canton herbal tea. I. asprella is readily contaminated by mildew because of rich nutrients. Aspergillus versicolor producing sterigmatocystin is one of the most common molds that contaminate foodstuffs and medicinal herbs. Previous study on the evaluation of fungal contamination on samples of I. asprella revealed that A. versicolor was the dominant contaminant. In this study, experiments based on response surface methodology combined with central composite design were carried out to determine the optimal storage conditions of I. asprella to minimize the contamination of sterigmatocystin. The herb, manually innoculated with A. versicolor, was stored under different temperatures (20⁻40 °C) and humidity (80⁻95%) for seven days. The effects of temperature and humidity were evaluated using total saponin, polysaccharide and the sterigmatocystin levels as quality indexes. The results showed that A. versicolor grew quickly and produced large amounts of sterigmatocystin on I. asprella, at humidity ranging from 85% to 90% and temperatures above 26 °C. Meanwhile, total saponin and polysaccharide amounts were reduced significantly. These findings suggested that I. asprella samples should be stored in an environment with humidity and temperature below 85% and 26 °C, respectively, to reduce A. versicolor growth and sterigmatocystin production.

[Simultaneous determination of six aflatoxins and sterigmatocystin in livestock and poultry tissues by QuEChERS-ultra high performance liquid chromatography-tandem mass spectrometry].

A method for the simultaneous determination of six aflatoxins (AFTs) and sterigmatocystin (SMC) in livestock and poultry tissues by QuEChERS-ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed and validated. The samples were extracted with 10 mL acetonitrile-water (84:16, v/v) after enzymatic hydrolysis with 20 µL ß-glucosidase. The extracts were cleaned with 1.0 g each of sodium chloride and anhydrous magnesium sulfate, and then degreased with 5 mL hexane. The residues were re-dissolved in 1 mL acetonitrile-water (80:20, v/v) containing 5 mmol/L ammonium acetate. Methanol and 5 mmol/L ammonium acetate were used as the mobile phases. Six AFTs and SMC were analyzed by multiple reaction monitoring in the positive ion mode. Matrix-matched calibration curves and external standards were used for the accurate quantification of the six AFTs and SMC. Good linear relationships were obtained, the correlation coefficients (R2) were greater than 0.99. The limits of detection (LODs, S/N=3) and the limits of quantification (LOQs, S/N=10) ranged from 0.007 to 0.30 µg/kg, and 0.02 to 0.91 µg/kg, respectively. The recoveries at different spiked levels were satisfactory, and ranged from 77.3% to 118.5%. This method is simple, easy and sensitive, and is suitable for the rapid determination of the six AFTs and SMC in different pork, pork liver and chicken samples.

Sterigmatocystin: A mycotoxin to be seriously considered.

Sterigmatocystin is a carcinogenic compound that affects several species of crops and several species of experimental animals. The sterigmatocystin biosynthetic pathway is the best known and most studied. The International Agency for Research on Cancer classifies sterigmatocystin in the Group 2B. Three groups of analytical methods to determine sterigmatocystin in food can be found: chromatographic, ELISA immunoassays and chemical sensors. In addition, sterigmatocystin is a precursor of aflatoxin B1 in those cases where cereals and/or food are contaminated with fungi capable of producing aflatoxins. Chemical structures of sterigmatocystin and aflatoxin B1 are similar. These mycotoxins are pathogens of animals and cereals, producing a major economic impact on biotechnology and agricultural and food industries. This review summarizes different aspects related to sterigmatocystin such as its biosynthesis, toxicological studies and analytical methods for its determination.

Development and in-house validation of a rapid and simple to use ELISA for the detection and measurement of the mycotoxin sterigmatocystin.

Sterigmatocystin (STG) is a highly toxic secondary fungal metabolite structurally closely related to the well-known carcinogenic aflatoxins. Its presence has been reported in grains and grain-based products as well as in other foodstuffs like nuts, green coffee beans, spices, beer and cheese. Due to the lack of suitable data on the occurrence of STG, in 2013, the European Food Safety Authority (EFSA) could not characterise its risk for human health and recommended that more data on STG in food and feed needed to be collected. In order to provide a new tool for the specific detection of STG, a competitive enzyme-linked immunosorbent assay (ELISA) was developed, optimised and validated in this study based on a sensitive monoclonal antibody specific to STG with no cross-reactivity with aflatoxins. The sample preparation method for rice, wheat and maize was based on a modified QuEChERS (quick, easy, cheap, effective, rugged and safe) approach. The assay was validated for the detection of STG in rice, wheat and maize in accordance with the guidelines for validation of semi-quantitative screening methods included in Commission Regulation (EU) 519/2014. The screening target concentration (STC) was set at 1.5 µg/kg. The cutoffs for rice, wheat and maize were 1.2, 1.2 and 1.3 µg/kg and the false suspected rates were 0.34, 1.15 and 0.78%, respectively. Good correlation was found between the results obtained by the STG ELISA and LC-MS/MS method for naturally contaminated rice samples. This validated method can be applied as a sensitive and high-throughput screening for the presence of STG in a range of agricultural commodities. Graphical abstract A new enzyme-linked immunosorbent assay based on an antibody specific to sterigmatocystin for the detection of this mycotoxin in corn, wheat and rice.