Identification Of Mycotoxins Research Articles

Identification of mycotoxins in yogurt samples using an optimized QuEChERS extraction and UHPLC-MS/MS detection.

Yogurt, a milk-derived product, is susceptible to mycotoxin contamination. While various methods have been developed for the analysis of dairy products, only a few have been specifically validated for yogurt. In addition, these methods are primarily focus on detecting aflatoxins and zearalenone. This study aimed to conduct a preliminary investigation into the presence of regulated, emerging, and modified mycotoxins in natural and oat yogurts available in the Spanish market. For this, a QuEChERS-based extraction method was optimized and then validated to detect and quantify 32 mycotoxins using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The method was in-house validated for the analysis of natural and oat yogurt in terms of linearity, matrix effect, sensitivity, accuracy, and precision. Satisfactory performance characteristics were achieved; for most of the analytes, LOQs were lower than 2ng/g, and recoveries ranged from 60 to 110% with a precision, expressed as the relative standard deviation of the recovery, lower than 15%. Subsequently, the validated method was applied to analyze commercial yogurt samples, revealing a notable incidence of beauvericin and enniatins, with some analogues found in up to 100% of the samples. Alternariol methyl ether was also frequently found, appearing in 50% of the samples. Additionally, the study identified regulated toxins such as fumonisins, ochratoxin A , and HT-2 toxin. These results provide new incidence data in yogurt, raising concerns about potential health risks for consumers.

Detection of alpha-amanitin encoding gene utilizing real-time PCR technique

Given the limited taxonomy of toxic fungi as well as the large number of mushroom species, identification of mycotoxins in mushroom samples can accelerate information gathering and progress, while increasing Enhanced testing accuracy of testing. In that situation, it is necessary to research methods to identify toxin-carrying genes in mushrooms, to help monitor and classify poisonous mushrooms, disseminate and increase people's awareness of food safety. done faster and more accurately. The research team succeeded in developing a real-time PCR method to detect the toxin alpha-amanitin gene on mushrooms with a detection limit of 101 copies/µL. The study has fully verified the correct parameters, 100% accuracy, and specificity.

Comparative Evaluation of Three Commercial Elisa Kits Used for the Detection of Aflatoxins B1, B2, G1, and G2 in Feedstuffs and Comparison with an HPLC Method.

Various analytical techniques for detecting mycotoxins have been developed in order to control their concentration in food and feed. Conventional analytical approaches for mycotoxin identification include thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), and gas chromatography (GC). Rapid methods for mycotoxin analysis are also becoming increasingly relevant. One of the most common rapid methods for determining these compounds is the enzyme-linked immunosorbent assay (ELISA). The current study aimed to compare three available ELISA kits for the detection and quantification of aflatoxins B1, B2, G1, and G2 in spiked feed samples at known quantities. All three ELISA kits were validated and showed good performance with high recovery rates and LOD and LOQ values lower than the MRL. The developed HPLC-FL method was validated for all the compounds determining the accuracy, precision, linearity, decision limit, and detection capability with fairly good results. Unknown feed samples (corn, silage, pellet, barley, wheat, soya, and sunflower) were also tested using the best ELISA kit and HPLC, and the results were compared. Both ELISA and HPLC were proven to be suitable methods for mycotoxin analysis. The analytical technique should be determined primarily by the availability and number of samples.

Raman Spectroscopy Enables Non-Invasive Identification of Mycotoxins p. Fusarium of Winter Wheat Seeds

Identification of specific mycotoxins p. Fusarium contained in infected winter wheat seeds can be achieved by visually recognizing their distinctive phenotypic species. The visual identification (ID) of species is subjective and usually requires significant taxonomic knowledge. Methods for the determination of various types of mycotoxins of the p. Fusarium are laborious and require the use of chemical invasive research methods. In this research, we investigate the possibility of using Raman spectroscopy (RS) as a tag-free, non-invasive and non-destructive analytical method for the rapid and accurate identification of p. Fusarium. Varieties of the r. Fusarium can produce mycotoxins that directly affect the DNA, RNA and chemical structure of infected seeds. Analysis of spectra by RS methods and chemometric analysis allows the identification of healthy, infected and contaminated seeds of winter wheat with varieties of mycotoxins p. Fusarium. Raman seed analysis provides accurate identification of p. Fusarium in 96% of samples. In addition, we present data on the identification of carbohydrates, proteins, fiber and other nutrients contaminated with p. Fusarium seeds obtained using spectroscopic signatures. These results demonstrate that RS enables rapid, accurate and non-invasive screening of seed phytosanitary status.

Mycotoxin Identification and In Silico Toxicity Assessment Prediction in Atlantic Salmon.

The present study aimed to identify mycotoxins in edible tissues of Atlantic salmon (Salmo salar) using liquid chromatography coupled to hybrid quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). After using a non-targeted screening approach and a home-made spectral library, 233 mycotoxins were analyzed. Moreover, the occurrence of mycotoxins in fish filets was evaluated, and their potential toxicity was predicted by in silico methods. According to the obtained results, forty mycotoxins were identified in analyzed salmon samples, the predominant mycotoxins being enniatins (also rugulosin and 17 ophiobolins), commonly found in cereals and their by-products. Thus, mycotoxin carry-over can occur from feed to organs and edible tissues of cultivated fish. Moreover, the toxicity of detected mycotoxins was predicted by the in silico webserver ProTox-II, highlighting that special attention must be paid to some less reported mycotoxins due to their toxic predicted properties.

Molecular detection of mycobiota and the associated mycotoxins in rice grains imported into Saudi Arabia

Rice is a staple food consumed across the world and its purity is essential to manage human health. However, fungal mycotoxins are one of the real challenges for maintaining the purity of rice intended for human consumption. The present study was designed to detect the fungal isolates present in the imported rice cultivars obtained from the local markets of the Kingdom of Saudi Arabia and the mycotoxins they produce using molecular approaches. The morphological analysis and PCR-based sequencing of ITS-rDNA specify that the isolates in the contaminated rice belonged to Aspergillus and Penicillium genera. The phylogenetic analysis based on sequences of ITS-rDNA regions of the isolates confirmed that the Aspergillus isolates were homological to A. flavus and A. nidulans. Whereas, the Penicillium isolates were homological to P. commune and P. chrysogenum. The identification of mycotoxins using LC-MS revealed that the isolates of the genus Aspergillus produced aflatoxin and ochratoxin. The isolates of Penicillium genus produced citreoviridin and citrinin. The levels of mycotoxins varied among each variety of the rice samples tested in total. Ochratoxin was produced at the least concentration (184.7 µg/kg) in the contaminated samples. Citrinin (23170.5 µg/kg) was produced at higher volumes in comparison to other mycotoxins. These levels of mycotoxin contamination are alarming and raise significant queries regarding the purity of the imported rice grains throughout the period of harvest and storage. Therefore, the development of practical control is required for consumer safety.

Mycotoxin Incidence in Some Fish Products: QuEChERS Methodology and Liquid Chromatography Linear Ion Trap Tandem Mass Spectrometry Approach.

The inclusion of vegetal raw materials in feed for fish farming has increased the risk of mycotoxin occurrence in feed, as well as in edible tissues from fish fed with contaminated feed, due to the carry-over to muscle portions. Therefore, the objective of this study was to evaluate the occurrence of 15 mycotoxins in processed fish products, which are commonly consumed, such as smoked salmon and trout, different types of sushi, and gula substitutes. A QuEChERS method was employed to perform the mycotoxin extraction from fish samples. For mycotoxin identification and quantitation, the selected technique was the liquid chromatography-tandem mass spectrometry linear ion trap (LC-MS/MS-LIT). Smoked fish and sushi samples results were negative regarding the presence of all 15 mycotoxins studied. In contrast, small amounts of fusarenon-X and enniatin B were found in gula substitute samples.

Mycotoxigenic potential of Alternaria alternata isolated from dragon fruit (Hylocereus undatus Haw.) using UHPLC-Qtof-MS

The occurrence of mycotoxins synthesized by spoilage fungi has been reported in various crops. Despite being a crop susceptible to attack by microorganisms, such as Alternaria alternata, there are no reports on the production of mycotoxins in Hylocereus undatus (Haw.). The objective of the present study was to identify mycotoxins produced by A. alternata previously isolated from Hylocereus undatus (Haw.) fruit in the postharvest phase, and A. alternata ATCC 46,582, in vitro and in vivo. The compounds were identified using liquid chromatography-mass spectrometry (LC–MS/MS). Alternariol (AOH), alternariol monomethyl ether (AME), tentoxin (TEN), altertoxins (ATX) I, II and III were detected in vitro for A. alternata, and ATX I and II were identified for the reference strain, A. alternata ATCC 46,582, which was used as a positive control. Mycotoxins characteristic of A. alternata were not detected in vivo in the fruit stored for 8 d, which is its estimated shelf life. The use of LC–MS/MS facilitated rapid and efficient mycotoxin identification, contributing significantly to the evaluation of food safety.

Identification of mycotoxins by UHPLC–QTOF MS in airborne fungi and fungi isolated from industrial paper and antique documents from the Archive of Bogotá

Mold deterioration of historical documents in archives and libraries is a frequent and complex phenomenon that may have important economic and cultural consequences. In addition, exposure to toxic fungal metabolites might produce health problems. In this work, samples of broths of fungal species isolated from the documentary material and from indoor environmental samples of the Archive of Bogotá have been analyzed to investigate the presence of mycotoxins. High resolution mass spectrometry made possible to search for a large number of mycotoxins, even without reference standards available at the laboratory. For this purpose, a screening strategy based on ultra-high pressure liquid chromatography coupled to quadrupole-time of flight mass spectrometry (UHPLC–QTOF MS) under MSE mode was applied. A customized home-made database containing elemental composition for around 600 mycotoxins was compiled. The presence of the (de)protonated molecule measured at its accurate mass was evaluated in the samples. When a peak was detected, collision induced dissociation fragments and characteristic isotopic ions were also evaluated and used for tentative identification, based on structure compatibility and comparison with literature data (if existing). Up to 44 mycotoxins were tentatively identified by UHPLC–QTOF MS. 34 of these tentative compounds were confirmed by subsequent analysis using a targeted LC–MS/MS method, supporting the strong potential of QTOF MS for identification/elucidation purposes. The presence of mycotoxins in these samples might help to reinforce safety measures for researchers and staff who work on reception, restoration and conservation of archival material, not only at the Archive of Bogotá but worldwide.

Family of Six, their Health and the Death of a 16 Month Old Male from Pulmonary Hemorrhage: Identification of Mycotoxins and Mold in the Home and Lungs, Liver and Brain of Deceased Infant.

Family of Six, their Health and the Death of a 16 Month Old Male from Pulmonary Hemorrhage: Identification of Mycotoxins and Mold in the Home and Lungs, Liver and Brain of Deceased Infant.

Rapid and Simultaneous In Situ Assessment of Aflatoxins and Stilbenes Using Silica Plate Imprinting Mass Spectrometry Imaging

A fast and direct combination of techniques for simultaneous mycotoxin and phytoalexin identification in peanut skin and kernel is described. Silica Plate Imprinting Laser Desorption/Ionization Mass Spectrometry Imaging (SPILDI-MSI) is a powerful technique that exhibits great advantages, such as solvent-free and matrix-free characteristics, as well as no sample preparation or separation steps. It also permits accurate identification of mycotoxins and phytoalexins with unique fingerprint profiles in just a few seconds. Results are expressed as chemical images of the 4 identified types of aflatoxins (B1, B2, G1 and G2) and a stilbenoid (resveratrol). Also, SPILDI-MSI allows the comparison between the spatial distribution of aflatoxins and resveratrol found in kernel and skin. This novel application has proven to be useful for instantaneous qualitative assessment of aflatoxins and stilbenoids both in the peanut skin and kernel and offers precise tracking of fungal contamination in nuts and other foodstuffs.

Combining the quick, easy, cheap, effective, rugged and safe approach and clean-up by immunoaffinity column for the analysis of 15 mycotoxins by isotope dilution liquid chromatography tandem mass spectrometry

Optimization and validation of a multi-mycotoxin method by LC–MS/MS is presented. The method covers the EU-regulated mycotoxins (aflatoxins, fumonisins, ochratoxin A, deoxynivalenol, zearalenone, T-2 and HT-2), as well as nivalenol and 3- and 15-acetyldeoxynivalenol for analysis of cereals, cocoa, oil, spices, infant formula, coffee and nuts. The proposed procedure combines two clean-up strategies: First, a generic preparation suitable for all mycotoxins based on the QuEChERS (for quick, easy, cheap, effective, rugged and safe) protocol. Second, a specific clean-up devoted to aflatoxins and ochratoxin A using immunoaffinity column (IAC) clean-up. Positive identification of mycotoxins in matrix was conducted according to the confirmation criteria defined in EU Commission Decision 2002/657/EC while quantification was performed by isotopic dilution using 13C-labeled mycotoxins as internal standards. Limits of quantification were at or below the maximum levels set in the EC/1886/2006 document for all mycotoxin/matrix combinations under regulation. In particular, the inclusion of an IAC step allowed achieving LOQs as low as 0.05 and 0.25μg/kg in cereals for aflatoxins and ochratoxin A, respectively. Other performance parameters like linearity [(r)2>0.99], recovery [71–118%], precision [(RSDr and RSDiR)<33%], and trueness [78–117%] were all compliant with the analytical requirements stipulated in the CEN/TR/16059 document. Method ruggedness was proved by a verification process conducted by another laboratory.

COMPARATIVE CHARACTERISTIC OF TRICHOTHECENE MYCOTOXIN COMPLEX OF FUSARIUM POAE (PECK) WOLLENW. THE STRAINS FROM DIFFERENT TROPHIC GROUPS

Aim. The main goal of our investigation was a comparative study of the ability to synthesize trichothecene toxins by 15 strains of Fusarium poae from different trophic groups: 5 soil strains, 5 plant pathogenic and 5 endophytic strains. Methods. The standard methods of isolation and identification of mycotoxins were used. Results. It was established that T-2 toxin was detected for 4 soil strains, 3 plant pathogenic and 2 endophytic strains. It was shown that its content was maximal for soil strains and reached 188.8±32.2 μg/l, was lower for plant pathogenic ones and the lowest for endophytic strains. The content of HT-2 toxin and T-2 tetraol was higher for soil strains than plant pathogenic and endophytic ones, and T-2 triol – higher for plant pathogens than soil strains. T-2 triol was observed only for one endophytic strain. Neosolaniol is produced by 3 soil and 2 plant pathogenic strains, but not synthesized by endophytes. Conclusions. Thus, it was established that endophytes F. poae produce significantly less trichothecene mycotoxins than the strains of this species from other trophic groups. This physiological peculiarity allows them apparently to be in a mutualism with the plants.

Development of a European system for identification of emerging mycotoxins in wheat supply chains

This paper describes the results of a research project aimed at the conceptual development of an identification system for emerging mycotoxins in European feed and food supply chains of wheat. Basic requirements for such a system were addressed, including the selection of indicators, locating information sources, development of underlying model, and investigation of stakeholders' needs. The selection and ranking of key indicators for identifying emerging mycotoxins was based on a literature review, followed by a structured expert judgment study. The expert study was based on the Delphi technique and used a panel of 43 European experts. The Delphi procedure resulted in a selected set of 12 key indicators for each of three relevant stages of the wheat chain (cultivation; transport and storage; processing). For wheat cultivation, these were: relative humidity/rainfall, crop rotation, temperature, tillage practice, water activity in the kernels, crop variety, harvest conditions, changes in fungal populations, fungicide use, plant health, regional infection pressure and food safety awareness. For the majority of the selected indicators, information sources were identified in specific European countries. However, most sources are not readily accessible and particular data is lacking. A theoretical concept for a model underlying the envisioned emerging mycotoxin identification system has been developed. The model links the key indicators and their information sources to aid and promote identification of emerging mycotoxins. It manages different types of information sources and levels of available information. The needs of various stakeholder groups regarding the system were investigated by means of two empirical elicitation sessions. The results showed that all stakeholders considered that such a system would be beneficial. One major challenge expected in establishing the system is stakeholder commitment to sharing data and information. This requires great efforts in improving communication and interaction between the key stakeholders.

Mycotoxin identification in moldy dwellings

The objective of this study was to assess the level of macrocyclic trichothecenes (MCT), one of the most potent mycotoxins, on wall surfaces, floor dust and air samples from moldy dwellings. The study was based on an index group comprising 15 flooded dwellings contaminated by Stachybotrys chartarum or Chaetomium and a control group comprising nine dwellings without molds on visual inspection and mold sampling. Three samples were collected from each dwelling: a sample from the moldy wall, using a swab, a floor dust sample on a 0.5 m(2) surface with a cloth and an air sample using a pump comprising a 1 microm Teflon filter. The MCT level was measured using a monoclonal, antibody-based ELISA test. Compared with measurements performed in index dwellings, higher MCT values were measured on floor dust samples from moldy dwellings (P = 0.02). Samples from wall surfaces demonstrated nearly significant differences (P = 0.06). No significant differences were observed for air samples (P = 0.15), but some samples showed increased MCT levels. Significant correlations were observed between the levels measured on wall surfaces, floor dust and air samples (P = 0.02 to 0.05). In contrast, no correlation was observed between moldy surface and MCT levels. In conclusion, this paper reports for the first time direct evidence for the presence of MCT in moldy dwellings.

Implementation of the Electronic Nose for the Identification of Mycotoxins in Durum Wheat (Triticum durum)

Implementation of the Electronic Nose for the Identification of Mycotoxins in Durum Wheat (Triticum durum)

Identification of mycotoxins in keratomycosis-derived Fusarium isolates by gas chromatography—mass spectrometry

The production of mycotoxins from Fusarium species has been demonstrated in isolates cultured from patients suffering from keratomycosis. The method employed a combination of thin-layer chromatography directly performed on gel plugs taken from the growth medium, cartridge column chromatography, silylation and gas chromatography on a non-polar stationary phase capillary column linked to mass spectrometry. The sensitivities of detection obtained for a signal-to-noise ratio of 33:1, were 200 pg for single stage GC-MS and 20 pg using tandem GC-MS-MS. Two mycotoxins, diacetoxyscirpenol and T-2 toxin were identified in three cultures.

Separation and Identification of Mycotoxins by Thin-Layer Chromatography/Fast Atom Bombardment Mass Spectrometry

The separation and identification of aflatoxins and trichothecenes in submicrogram quantity by thin-layer chromatography/fast atom bombardment (TLC/ FAB) mass spectrometry is reported. Separated TLC spots (Rf values given) were put directly to an FAB probe tip along with a glycerol matrix. Two dimensional TLC in the solvent systems used could separate a mixture of aflatoxins (M1, M2, B2, G1, G2) and trichothecenes (HT-2, T-2). FAB mass spectral data are characteristic in identifying individual components.

Identification of mycotoxins produced by species of Fusarium and Stachybotrys obtained from Eastern Europe.

Isolates of Fusarium and Stachybotrys spp. and crude extracts from these fungi were obtained from Hungary and the U.S.S.R. and used for the evaluation of the mycotoxins they produced. The cultures were grown on millet and oats and extracted in Budapest, Hungary (Veterinary Medical Research Institute) and chemically analyzed at the University of Minnesota using thin-layer chromatography (TLC), gas-liquid chromatography (GLC), gas chromatograph-mass spectrometry (GC-MS), and the rat skin bioassay. Zearalenone was found in most of the Fusarium cultures, T-2 toxin, neosolaniol, T-2 tetraol, and HT-2 toxin were found in extracts of Fusarium poae and F. sporotrichioies. A special effort was made to isolate the steroid-like toxins reported in the early Russian literature as sporofusarin and poaefusarin. None of the extracts from the Fusarium species yielded poaefusarin or sporofusarin when analyzed by our chemical methods or by those of L.E. Olifson, S.M. Kenina, and V.L. Kartashova, 1972. We therefore accounted for the toxicity of the Fusarium extracts as due to the 12,13,epoxytrichothecenes. One culture of Stachybotrys alternans yielded a macrocyclic ester of 12,13-epoxytrichothecene which, upon hydrolysis, yielded verrucarol; a steroid-like molecule (SB-3) was also isolated. The former had skin-irritant activity but SB-3 did not; the latter exhibited cardiac activity on the heart of the cockroach.