Rapid Detection of Mycotoxin Contamination

A special issue of Toxins (ISSN 2072-6651). This special issue belongs to the section "Mycotoxins".

Deadline for manuscript submissions: closed (31 December 2020) | Viewed by 57694

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Guest Editor
Agro-Environmental Research Institute, National Agricultural Research and Innovation Centre, Herman O. u. 15, H-1022 Budapest, Hungary
Interests: environmental and food safety; organic microcontaminants (pesticide residues and mycotoxins); environmental analysis; agricultural ecotoxicology; genetic safety
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Special Issue Information

Dear Colleagues,

Mycotoxin contamination in crops, and the subsequent mycotoxin contamination in food and feed is currently a major concern in environmental and food safety, affecting both crop production and animal husbandry. In turn, the rapid detection of mycotoxin levels in food and feed, as well as in other biological and environmental matrices, is of key importance both in mycotoxin monitoring and exposure assessment.

Mycotoxin occurrence in produce is mostly as a result of improper harvest or storage conditions that favour the emergence of toxinogenic fungi (e.g., Fusarium, Penicillium, Aspergillus, and other species). Target mycotoxins include the most hazardous aflatoxins, trichothecenes (e.g., T-2, deoxynivalenol), resorcilactones (e.g., zearalenone), fumonisins, and ochratoxins, as well as recently identified compounds (e.g., sterigmatocystin, moniliformin, and others). The meteorological conditions prior to harvest strongly affect fungal growth and mycotoxin production; moreover, climate change also exerts its impact, as toxinogenic fungal strains may now emerge at climatic zones where they could not colonise before.

Our Special Issue of Toxins aims to summarise the importance of mycotoxin detection in various matrices by reporting diverse aspects, hopefully covering a wide range of application, including (but not limited to) the following:

- monitoring the occurrence of mycotoxins in crops and produce, as related to meteorological conditions, including the assessment of the potential effects of climate change trends on mycotoxin occurrence;

- a particular issue related to the abovementioned point is the general and repeatedly refuted allegation of ecological farming, of being a source of mycotoxin contamination because of the prohibition of the use of synthetic fungicides; therefore, the submission of comparative monitoring studies of mycotoxins in conventional and ecological agriculture are welcome;

- decomposition of mycotoxins in biological matrices, because of the effects of natural or artificially accelerated enzymatic conditions;

- effect-based monitoring of mycotoxins in affected animals, as well as veterinary mycotoxin analyses;

- assessment of mycotoxin decontamination methods aiming to suppress emerging mycotoxin poisoning;

- novel or inventive methods of mycotoxin analysis, including chromatography, immunoassay, molecular biology, sensorics, and other means, including novel sample preparation methods (e.g., QuEChERS and immunoaffinity pre-purification);

- methods of toxicological or ecotoxicological assessment, including cytotoxicity, genotoxicity, mutagenicity, and endocrine disruption, combined with chemical analysis.

Prof. András Székács
Guest Editor

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Keywords

  • mycotoxin analysis
  • monitoring
  • decomposition/metabolism
  • decontamination
  • instrumental and immunoanalysis
  • sensorics
  • cytotoxicity
  • genotoxicity
  • mutagenicity
  • endocrine disruption

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Published Papers (14 papers)

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Editorial

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5 pages, 263 KiB  
Editorial
Mycotoxins as Emerging Contaminants. Introduction to the Special Issue “Rapid Detection of Mycotoxin Contamination”
by András Székács
Toxins 2021, 13(7), 475; https://doi.org/10.3390/toxins13070475 - 9 Jul 2021
Cited by 8 | Viewed by 2709
Abstract
Concerns for human and environmental health regarding mycotoxins are predominantly raised in connection with their occurrence in food and feed (especially in grains) [...] Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)

Research

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22 pages, 684 KiB  
Article
Improved Sample Selection and Preparation Methods for Sampling Plans Used to Facilitate Rapid and Reliable Estimation of Aflatoxin in Chicken Feed
by James Kibugu, Raymond Mdachi, Leonard Munga, David Mburu, Thomas Whitaker, Thu P. Huynh, Delia Grace and Johanna F. Lindahl
Toxins 2021, 13(3), 216; https://doi.org/10.3390/toxins13030216 - 16 Mar 2021
Cited by 3 | Viewed by 3956
Abstract
Aflatoxin B1 (AFB1), a toxic fungal metabolite associated with human and animal diseases, is a natural contaminant encountered in agricultural commodities, food and feed. Heterogeneity of AFB1 makes risk estimation a challenge. To overcome this, novel sample selection, preparation and extraction steps were [...] Read more.
Aflatoxin B1 (AFB1), a toxic fungal metabolite associated with human and animal diseases, is a natural contaminant encountered in agricultural commodities, food and feed. Heterogeneity of AFB1 makes risk estimation a challenge. To overcome this, novel sample selection, preparation and extraction steps were designed for representative sampling of chicken feed. Accuracy, precision, limits of detection and quantification, linearity, robustness and ruggedness were used as performance criteria to validate this modification and Horwitz function for evaluating precision. A modified sampling protocol that ensured representativeness is documented, including sample selection, sampling tools, random procedures, minimum size of field-collected aggregate samples (primary sampling), procedures for mass reduction to 2 kg laboratory (secondary sampling), 25 g test portion (tertiary sampling) and 1.3 g analytical samples (quaternary sampling). The improved coning and quartering procedure described herein (for secondary and tertiary sampling) has acceptable precision, with a Horwitz ratio (HorRat = 0.3) suitable for splitting of 25 g feed aliquots from laboratory samples (tertiary sampling). The water slurring innovation (quaternary sampling) increased aflatoxin extraction efficiency to 95.1% through reduction of both bias (−4.95) and variability of recovery (1.2–1.4) and improved both intra-laboratory precision (HorRat = 1.2–1.5) and within-laboratory reproducibility (HorRat = 0.9–1.3). Optimal extraction conditions are documented. The improved procedure showed satisfactory performance, good field applicability and reduced sample analysis turnaround time. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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17 pages, 2530 KiB  
Article
Development of an Immunofluorescence Assay Module for Determination of the Mycotoxin Zearalenone in Water
by Borbála Gémes, Eszter Takács, Patrik Gádoros, Attila Barócsi, László Kocsányi, Sándor Lenk, Attila Csákányi, Szabolcs Kautny, László Domján, Gábor Szarvas, Nóra Adányi, Alexei Nabok, Mária Mörtl and András Székács
Toxins 2021, 13(3), 182; https://doi.org/10.3390/toxins13030182 - 2 Mar 2021
Cited by 11 | Viewed by 3646
Abstract
Project Aquafluosense is designed to develop prototypes for a fluorescence-based instrumentation setup for in situ measurements of several characteristic parameters of water quality. In the scope of the project an enzyme-linked fluorescent immunoassay (ELFIA) method has been developed for the detection of several [...] Read more.
Project Aquafluosense is designed to develop prototypes for a fluorescence-based instrumentation setup for in situ measurements of several characteristic parameters of water quality. In the scope of the project an enzyme-linked fluorescent immunoassay (ELFIA) method has been developed for the detection of several environmental xenobiotics, including mycotoxin zearalenone (ZON). ZON, produced by several plant pathogenic Fusarium species, has recently been identified as an emerging pollutant in surface water, presenting a hazard to aquatic ecosystems. Due to its physico-chemical properties, detection of ZON at low concentrations in surface water is a challenging task. The 96-well microplate-based fluorescence instrument is capable of detecting ZON in the concentration range of 0.09–400 ng/mL. The sensitivity and accuracy of the analytical method has been demonstrated by a comparative assessment with detection by high-performance liquid chromatography and by total internal reflection ellipsometry. The limit of detection of the method, 0.09 ng/mL, falls in the low range compared to the other reported immunoassays, but the main advantage of this ELFIA method is its efficacy in combined in situ applications for determination of various important water quality parameters detectable by induced fluorimerty—e.g., total organic carbon content, algal density or the level of other organic micropollutants detectable by immunofluorimetry. In addition, the immunofluorescence module can readily be expanded to other target analytes if proper antibodies are available for detection. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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12 pages, 2662 KiB  
Article
An Optical Planar Waveguide-Based Immunosensors for Determination of Fusarium Mycotoxin Zearalenone
by Alexei Nabok, Ali Madlool Al-Jawdah, Borbála Gémes, Eszter Takács and András Székács
Toxins 2021, 13(2), 89; https://doi.org/10.3390/toxins13020089 - 25 Jan 2021
Cited by 15 | Viewed by 3053
Abstract
A planar waveguide (PW) immunosensor working as a polarisation interferometer was developed for the detection of mycotoxin zearalenone (ZON). The main element of the sensor is an optical waveguide consisting of a thin silicon nitride layer between two thicker silicon dioxide layers. A [...] Read more.
A planar waveguide (PW) immunosensor working as a polarisation interferometer was developed for the detection of mycotoxin zearalenone (ZON). The main element of the sensor is an optical waveguide consisting of a thin silicon nitride layer between two thicker silicon dioxide layers. A combination of a narrow waveguiding core made by photolithography with an advanced optical set-up providing a coupling of circular polarised light into the PW via its slanted edge allowed the realization of a novel sensing principle by detection of the phase shift between the p- and s-components of polarised light propagating through the PW. As the p-component is sensitive to refractive index changes at the waveguide interface, molecular events between the sensor surface and the contacting sample solution can be detected. To detect ZON concentrations in the sample solution, ZON-specific antibodies were immobilised on the waveguide via an electrostatically deposited polyelectrolyte layer, and protein A was adsorbed on it. Refractive index changes on the surface due to the binding of ZON molecules to the anchored antibodies were detected in a concentration-dependent manner up to 1000 ng/mL of ZON, allowing a limit of detection of 0.01 ng/mL. Structurally unrelated mycotoxins such as aflatoxin B1 or ochratoxin A did not exert observable cross-reactivity. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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15 pages, 1939 KiB  
Article
Direct and Competitive Optical Grating Immunosensors for Determination of Fusarium Mycotoxin Zearalenone
by Inna Székács, Nóra Adányi, István Szendrő and András Székács
Toxins 2021, 13(1), 43; https://doi.org/10.3390/toxins13010043 - 8 Jan 2021
Cited by 12 | Viewed by 3074
Abstract
Novel optical waveguide lightmode spectroscopy (OWLS)-based immunosensor formats were developed for label-free detection of Fusarium mycotoxin zearalenone (ZON). To achieve low limits of detection (LODs), both immobilised antibody-based (direct) and immobilised antigen-based (competitive) assay setups were applied. Immunoreagents were immobilised on epoxy-, amino-, [...] Read more.
Novel optical waveguide lightmode spectroscopy (OWLS)-based immunosensor formats were developed for label-free detection of Fusarium mycotoxin zearalenone (ZON). To achieve low limits of detection (LODs), both immobilised antibody-based (direct) and immobilised antigen-based (competitive) assay setups were applied. Immunoreagents were immobilised on epoxy-, amino-, and carboxyl-functionalised sensor surfaces, and by optimising the immobilisation methods, standard sigmoid curves were obtained in both sensor formats. An outstanding LOD of 0.002 pg/mL was obtained for ZON in the competitive immunosensor setup with a dynamic detection range between 0.01 and 1 pg/mL ZON concentrations, depending on the covalent immobilisation method applied. This corresponds to a five orders of magnitude improvement in detectability of ZON relative to the previously developed enzyme-linked immonosorbent assay (ELISA) method. The selectivity of the immunosensor for ZON was demonstrated with structural analogues (α-zearalenol, α-zearalanol, and β-zearalanol) and structurally unrelated mycotoxins. The method was found to be applicable in maize extract using acetonitrile as the organic solvent, upon a dilution rate of 1:10,000 in buffer. Thus, the OWLS immunosensor method developed appears to be suitable for the quantitative determination of ZON in aqueous medium. The new technique can widen the range of sensoric detection methods of ZON for surveys in food and environmental safety assessment. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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12 pages, 3427 KiB  
Article
Aflatoxin B1 and Sterigmatocystin Binding Potential of Non-Lactobacillus LAB Strains
by Ildikó Bata-Vidács, Judit Kosztik, Mária Mörtl, András Székács and József Kukolya
Toxins 2020, 12(12), 799; https://doi.org/10.3390/toxins12120799 - 14 Dec 2020
Cited by 12 | Viewed by 2742
Abstract
Research on the ability of lactic acid bacteria (LAB) to bind aflatoxin B1 (AFB1) has mostly been focusing on lactobacilli and bifidobacteria. In this study, the AFB1 binding capacities of 20 Enterococcus strains belonging to E. casseliflavus, E. faecalis, E. faecium [...] Read more.
Research on the ability of lactic acid bacteria (LAB) to bind aflatoxin B1 (AFB1) has mostly been focusing on lactobacilli and bifidobacteria. In this study, the AFB1 binding capacities of 20 Enterococcus strains belonging to E. casseliflavus, E. faecalis, E. faecium, E. hirae, E. lactis, and E. mundtii, 24 Pediococcus strains belonging to species P. acidilactici, P. lolii, P. pentosaceus, and P. stilesii, one strain of Lactococcus formosensis and L.garviae, and 3 strains of Weissella soli were investigated in MRS broth at 37 °C at 0.2 µg/mL mycotoxin concentration. According to our results, among non-lactobacilli LAB, the genera with the best AFB1 binding abilities were genus Pediococcus, with a maximum binding percentage of 7.6% by P. acidilactici OR83, followed by genus Lactococcus. For AFB1 bio-detoxification purposes, beside lactobacilli, pediococci can also be chosen, but it is important to select a strain with better binding properties than the average value of its genus. Five Pediococcus strains have been selected to compare their sterigmatocystin (ST) binding abilities to AFB1 binding, and a 2–3-fold difference was obtained similar to previous findings for lactobacilli. The best strain was P. acidilactici OR83 with 18% ST binding capacity. This is the first report on ST binding capabilities of non-Lactobacillus LAB strains. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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16 pages, 7381 KiB  
Article
Aflatoxin B1 and Sterigmatocystin Binding Potential of Lactobacilli
by Judit Kosztik, Mária Mörtl, András Székács, József Kukolya and Ildikó Bata-Vidács
Toxins 2020, 12(12), 756; https://doi.org/10.3390/toxins12120756 - 30 Nov 2020
Cited by 15 | Viewed by 2879
Abstract
Due to global climate change, mould strains causing problems with their mycotoxin production in the tropical–subtropical climate zone have also appeared in countries belonging to the temperate zone. Biodetoxification of crops and raw materials for food and feed industries including the aflatoxin B1 [...] Read more.
Due to global climate change, mould strains causing problems with their mycotoxin production in the tropical–subtropical climate zone have also appeared in countries belonging to the temperate zone. Biodetoxification of crops and raw materials for food and feed industries including the aflatoxin B1 (AFB1) binding abilities of lactobacilli is of growing interest. Despite the massive quantities of papers dealing with AFB1-binding of lactobacilli, there are no data for microbial binding of the structurally similar mycotoxin sterigmatocystin (ST). In addition, previous works focused on the detection of AFB1 in extracts, while in this case, analytical determination was necessary for the microbial biomass as well. To test binding capacities, a rapid instrumental analytical method using high-performance liquid chromatography was developed and applied for measurement of AFB1 and ST in the biomass of the cultured bacteria and its supernatant, containing the mycotoxin fraction bound by the bacteria and the fraction that remained unbound, respectively. For our AFB1 and ST adsorption studies, 80 strains of the genus Lactobacillus were selected. Broths containing 0.2 µg/mL AFB1and ST were inoculated with the Lactobacillus test strains. Before screening the strains for binding capacities, optimisation of the experiment parameters was carried out. Mycotoxin binding was detectable from a germ count of 107 cells/mL. By studying the incubation time of the cells with the mycotoxins needed for mycotoxin-binding, co-incubation for 10 min was found sufficient. The presence of mycotoxins did not affect the growth of bacterial strains. Three strains of L. plantarum had the best AFB1 adsorption capacities, binding nearly 10% of the mycotoxin present, and in the case of ST, the degree of binding was over 20%. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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12 pages, 2516 KiB  
Article
Contamination of Zearalenone from China in 2019 by a Visual and Digitized Immunochromatographic Assay
by Xia Hong, Yuhao Mao, Chuqin Yang, Zhenjiang Liu, Ming Li and Daolin Du
Toxins 2020, 12(8), 521; https://doi.org/10.3390/toxins12080521 - 14 Aug 2020
Cited by 19 | Viewed by 3365
Abstract
Zearalenone (ZEN) is a prevalent mycotoxin that needs intensive monitoring. A semi-quantitative and quantitative immunochromatographic assay (ICA) was assembled for investigating ZEN contamination in 187 samples of cereal and their products from China in 2019. The semi-quantitative detection model had a limit of [...] Read more.
Zearalenone (ZEN) is a prevalent mycotoxin that needs intensive monitoring. A semi-quantitative and quantitative immunochromatographic assay (ICA) was assembled for investigating ZEN contamination in 187 samples of cereal and their products from China in 2019. The semi-quantitative detection model had a limit of detection (LOD) of 0.50 ng/mL with visual judgment and could be completely inhibited within 5 min at 3.0 ng/mL ZEN. The quantitative detection model had a lower LOD of 0.25 ng/mL, and ZEN could be accurately and digitally detected from 0.25–4.0 ng/mL. The ICA method had a high sensitivity, specificity, and accuracy for on-site ZEN detection. For investigation of the authentic samples, the ZEN-positive rate was 62.6%, and the ZEN-positive levels ranged from 2.7 to 867.0 ng/g, with an average ZEN-positive level being 85.0 ng/g. Of the ZEN-positive samples, 6.0% exceeded the values of the limit levels. The ZEN-positive samples were confirmed to be highly correlated using LC-MS/MS (R2 = 0.9794). This study could provide an efficiency and accuracy approach for ZEN in order to achieve visual and digitized on-site investigation. This significant information about the ZEN contamination levels might contribute to monitoring mycotoxin occurrence and for ensuring food safety. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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14 pages, 740 KiB  
Article
Development of an Improved Method of Sample Extraction and Quantitation of Multi-Mycotoxin in Feed by LC-MS/MS
by Bahar Nakhjavan, Nighat Sami Ahmed and Maryam Khosravifard
Toxins 2020, 12(7), 462; https://doi.org/10.3390/toxins12070462 - 19 Jul 2020
Cited by 35 | Viewed by 6011
Abstract
A multi-mycotoxin chromatographic method was developed and validated for the simultaneous quantitation of aflatoxins (AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), zearalenone (ZON), deoxynivalenol (DON), nivalenol (NIV), diacetoxyscirpenol (DAS), fumonisins (FB1, FB2 and FB3), T-2 toxin (T-2) and HT-2 toxin (HT-2) in [...] Read more.
A multi-mycotoxin chromatographic method was developed and validated for the simultaneous quantitation of aflatoxins (AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), zearalenone (ZON), deoxynivalenol (DON), nivalenol (NIV), diacetoxyscirpenol (DAS), fumonisins (FB1, FB2 and FB3), T-2 toxin (T-2) and HT-2 toxin (HT-2) in feed. The three most popular sample preparation techniques for determination of mycotoxins have been evaluated, and the method with highest recoveries was selected and optimized. This modified QuEChERS (quick, easy, cheap, effective, rugged and safe) approach was based on the extraction with acetonitrile, salting-out and cleanup with lipid removal. A reconstitution process in methanol/water was used to improve the MS responses and then the extracts were analyzed by LC-MS/MS. In this method, the recovery range is 70–100% for DON, DAS, FB1, FB2, FB3, HT-2, T-2, OTA, ZON, AFG1, AFG2, AFB1 and AFB2 and 55% for NIV in the spike range of 2–80 µg/kg. Method robustness was determined with acceptable z-scores in proficiency tests and validation experiments. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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13 pages, 3365 KiB  
Article
A Non-Enzyme and Non-Label Sensitive Fluorescent Aptasensor Based on Simulation-Assisted and Target-Triggered Hairpin Probe Self-Assembly for Ochratoxin a Detection
by Mengyao Qian, Wenxiao Hu, Luhui Wang, Yue Wang and Yafei Dong
Toxins 2020, 12(6), 376; https://doi.org/10.3390/toxins12060376 - 6 Jun 2020
Cited by 16 | Viewed by 3693
Abstract
The monitoring and control of mycotoxins has caused widespread concern due to their adverse effects on human health. In this research, a simple, sensitive and non-label fluorescent aptasensor has been reported for mycotoxin ochratoxin A (OTA) detection based on high selectivity of aptamers [...] Read more.
The monitoring and control of mycotoxins has caused widespread concern due to their adverse effects on human health. In this research, a simple, sensitive and non-label fluorescent aptasensor has been reported for mycotoxin ochratoxin A (OTA) detection based on high selectivity of aptamers and amplification of non-enzyme hybridization chain reaction (HCR). After the introduction of OTA, the aptamer portion of hairpin probe H1 will combine with OTA to form OTA-aptamer complexes. Subsequently, the remainder of the opened H1 will act as an initiator for the HCR between the two hairpin probes, causing H1 and H2 to be sequentially opened and assembled into continuous DNA duplexes embedded with numerous G-quadruplexes, leading to a significant enhancement in fluorescence signal after binding with N-methyl-mesoporphyrin IX (NMM). The proposed sensing strategy can detect OTA with concentration as low as 4.9 pM. Besides, satisfactory results have also been obtained in the tests of actual samples. More importantly, the thermodynamic properties of nucleic acid chains in the monitoring platform were analyzed and the reaction processes and conditions were simulated before carrying out biological experiments, which theoretically proved the feasibility and simplified subsequent experimental operations. Therefore, the proposed method possess a certain application value in terms of monitoring mycotoxins in food samples and improving the quality control of food security. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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15 pages, 2163 KiB  
Article
Simultaneous Determination of Deoxynivalenol, Its Modified Forms, Nivalenol and Fusarenone-X in Feedstuffs by the Liquid Chromatography–Tandem Mass Spectrometry Method
by Łukasz Panasiuk, Piotr Jedziniak, Katarzyna Pietruszka and Andrzej Posyniak
Toxins 2020, 12(6), 362; https://doi.org/10.3390/toxins12060362 - 1 Jun 2020
Cited by 12 | Viewed by 3540
Abstract
A liquid chromatography-tandem mass spectrometry method was developed for simultaneous determination of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3Ac-DON), 15-acetyldeoxynivalenol (15Ac-DON), DON-3-glucoside (DON-3Glc) nivalenol and fusarenone-X in feedstuffs. Different techniques of sample preparation were tested: solid-liquid-extraction, QuEChERS, solid phase extraction with OASIS HLB columns or immunoaffinity [...] Read more.
A liquid chromatography-tandem mass spectrometry method was developed for simultaneous determination of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3Ac-DON), 15-acetyldeoxynivalenol (15Ac-DON), DON-3-glucoside (DON-3Glc) nivalenol and fusarenone-X in feedstuffs. Different techniques of sample preparation were tested: solid-liquid-extraction, QuEChERS, solid phase extraction with OASIS HLB columns or immunoaffinity columns and a Mycosep 225 Trich column. None of the six immunoaffinity columns tested showed cross-reactivity to all of the mycotoxins. Surprisingly, the results show that if the immunoaffinity columns bound 3Ac-DON, then they did not bind 15Ac-DON. The most efficient sample preparation was achieved with a Mycosep 225 Trich column clean-up. The chromatography was optimised to obtain full separation of all analytes (including 3Ac-DON and 15Ac-DON isomeric form). The validation results show the relative standard deviations for repeatability and reproducibility varied from 4% to 24%. The apparent recovery ranged between 92% and 97%, and the limit of quantification described a 1.30 to 50 µg/kg range. The method trueness was satisfactory, as assessed by a proficiency test and analysis of reference material. A total of 99 feed samples were analysed by the developed method, revealing the presence of DON and DON-3Glc in 85% and 86% of examined animal feeds, respectively at concentrations between 1.70 and 1709 µg/kg. The ratios DON-3Glc to DON in the surveyed feedstuffs were from a low of 3% to high of 59%. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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15 pages, 2832 KiB  
Article
Sensitive Aflatoxin B1 Detection Using Nanoparticle-Based Competitive Magnetic Immunodetection
by Jan Pietschmann, Holger Spiegel, Hans-Joachim Krause, Stefan Schillberg and Florian Schröper
Toxins 2020, 12(5), 337; https://doi.org/10.3390/toxins12050337 - 20 May 2020
Cited by 26 | Viewed by 5214
Abstract
Food and crop contaminations with mycotoxins are a severe health risk for consumers and cause high economic losses worldwide. Currently, different chromatographic- and immuno-based methods are used to detect mycotoxins within different sample matrices. There is a need for novel, highly sensitive detection [...] Read more.
Food and crop contaminations with mycotoxins are a severe health risk for consumers and cause high economic losses worldwide. Currently, different chromatographic- and immuno-based methods are used to detect mycotoxins within different sample matrices. There is a need for novel, highly sensitive detection technologies that avoid time-consuming procedures and expensive laboratory equipment but still provide sufficient sensitivity to achieve the mandated detection limit for mycotoxin content. Here we describe a novel, highly sensitive, and portable aflatoxin B1 detection approach using competitive magnetic immunodetection (cMID). As a reference method, a competitive ELISA optimized by checkerboard titration was established. For the novel cMID procedure, immunofiltration columns, coated with aflatoxin B1-BSA conjugate were used for competitive enrichment of biotinylated aflatoxin B1-specific antibodies. Subsequently, magnetic particles functionalized with streptavidin can be applied to magnetically label retained antibodies. By means of frequency mixing technology, particles were detected and quantified corresponding to the aflatoxin content in the sample. After the optimization of assay conditions, we successfully demonstrated the new competitive magnetic detection approach with a comparable detection limit of 1.1 ng aflatoxin B1 per mL sample to the cELISA reference method. Our results indicate that the cMID is a promising method reducing the risks of processing contaminated commodities. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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11 pages, 1898 KiB  
Article
A Liquid Chromatographic Method for Rapid and Sensitive Analysis of Aflatoxins in Laboratory Fungal Cultures
by Ahmad F. Alshannaq and Jae-Hyuk Yu
Toxins 2020, 12(2), 93; https://doi.org/10.3390/toxins12020093 - 30 Jan 2020
Cited by 17 | Viewed by 5503
Abstract
Culture methods supplemented with high-performance liquid chromatography (HPLC) technique provide a rapid and simple tool for detecting levels of aflatoxins (AFs) produced by fungi. This study presents a robust method for simultaneous quantification of aflatoxin (AF) B1, B2, G1, and G2 levels in [...] Read more.
Culture methods supplemented with high-performance liquid chromatography (HPLC) technique provide a rapid and simple tool for detecting levels of aflatoxins (AFs) produced by fungi. This study presents a robust method for simultaneous quantification of aflatoxin (AF) B1, B2, G1, and G2 levels in several fungal cultivation states: submerged shake culture, liquid slant culture, and solid-state culture. The recovery of the method was evaluated by spiking a mixture of AFs at several concentrations to the test medium. The applicability of the method was evaluated by using aflatoxigenic and non-aflatoxigenic Aspergilli. A HPLC coupled with the diode array (DAD) and fluorescence (FLD) detectors was used to determine the presence and amounts of AFs. Both detectors showed high sensitivity in detecting spiked AFs or AFs produced in situ by toxigenic fungi. Our methods showed 76%–88% recovery from medium spiked with 2.5, 10, 50, 100, and 500 ng/mL AFs. The limit of quantification (LOQ) for AFs were 2.5 to 5.0 ng/mL with DAD and 0.025 to 2.5 ng/mL with FLD. In this work, we described in detail a protocol, which can be considered the foremost and only verified method, to extract, detect, and quantify AFs employing both aflatoxigenic and non-toxigenic Aspergilli. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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Review

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35 pages, 3620 KiB  
Review
Advances in Colorimetric Strategies for Mycotoxins Detection: Toward Rapid Industrial Monitoring
by Marjan Majdinasab, Sondes Ben Aissa and Jean Louis Marty
Toxins 2021, 13(1), 13; https://doi.org/10.3390/toxins13010013 - 24 Dec 2020
Cited by 30 | Viewed by 5403
Abstract
Mycotoxins contamination is a global public health concern. Therefore, highly sensitive and selective techniques are needed for their on-site monitoring. Several approaches are conceivable for mycotoxins analysis, among which colorimetric methods are the most attractive for commercialization purposes thanks to their visual read-out, [...] Read more.
Mycotoxins contamination is a global public health concern. Therefore, highly sensitive and selective techniques are needed for their on-site monitoring. Several approaches are conceivable for mycotoxins analysis, among which colorimetric methods are the most attractive for commercialization purposes thanks to their visual read-out, easy operation, cost-effectiveness, and rapid response. This review covers the latest achievements in the last five years for the development of colorimetric methods specific to mycotoxins analysis, with a particular emphasis on their potential for large-scale applications in food industries. Gathering all types of (bio)receptors, main colorimetric methods are critically discussed, including enzyme-linked assays, lateral flow-assays, microfluidic devices, and homogenous in-solution strategies. This special focus on colorimetry as a versatile transduction method for mycotoxins analysis is comprehensively reviewed for the first time. Full article
(This article belongs to the Special Issue Rapid Detection of Mycotoxin Contamination)
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