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Analytical Methods for Mycotoxin Analysis
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Analytical Methods for Mycotoxin Analysis

A special issue of Molecules (ISSN 1420-3049). This special issue belongs to the section "Analytical Chemistry".

Deadline for manuscript submissions: closed (30 November 2020) | Viewed by 32478

Special Issue Editors

Dr. Terenzio Bertuzzi

E-Mail Website
Guest Editor
Department of Animal Science, Food and Nutrition, Faculty of Agricultural, Food and Environmental Sciences, UCSC, Via Emilia Parmense, 84, 29122 Piacenza, Italy
Interests: food safety; mycotoxin
Special Issues, Collections and Topics in MDPI journals
Dr. Chiara Lanzanova

E-Mail Website
Guest Editor
Council for Agricultural Research and Economics (CREA), Research Centre for Cereal and Industrial Crops, Via Stezzano, 24, 24126 Bergamo, Italy
Interests: fungal pathogens; mycotoxins; crop protection
Special Issues, Collections and Topics in MDPI journals
Dr. Sabrina Locatelli

E-Mail Website
Guest Editor
Council for Agricultural Research and Economics (CREA), Research Centre for Cereal and Industrial Crops, Via Stezzano, 24, 24126 Bergamo, Italy
Interests: mycotoxins; crops; safety
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Mycotoxins are secondary metabolites produced by fungi of different species, mainly Aspergillus, Fusarium, Penicillium, and Alternaria, which can contaminate food and feed with toxic effects for humans and animals. They represent the most important class of chemical hazards recorded in the European Rapid Alert System on Feed and Food (RASFF). In 2017, 43.4% of the alerts concerning a potential threat to human health from chemical contaminants in food products, mainly cereals and nuts, were due to mycotoxins.

Regulations around the world have established maximum levels for different mycotoxins in foodstuffs, including aflatoxins B1, B2, G1, G2, and M1, ochratoxin A, fumonisins B1 and B2, deoxynivalenol, zearalenone, HT-2 and T-2 toxins, patulin, citrinin, and ergot alkaloids. In addition to these “known and legislatively regulated” mycotoxins, there are other “emerging mycotoxins” that have been considered as relevant since evidence of their incidence in food and feed is rapidly increasing. This group includes Alternaria toxins, such as alternariol, alternariol monomethyl ether, and tenuazonic acid, Aspergillus toxins, such as sterigmatocystin, Fusarium toxins, such as moniliformin, enniantins and beauvericin, phomopsins, and others.

Therefore, it is necessary to develop analytical methods for an accurate determination of mycotoxins in different food matrices and feeds, achieving both the requirement of assuring food safety along the production chain and the development of advanced instrumental techniques to identify new emerging and masked mycotoxins. To obtain a continuous monitoring of these hazardous compounds in raw materials and final products along the production food chain, rapid, cheap, and easy-to-operate analytical methods are generally used; on the other hand, innovative approaches and advanced instrumental techniques, such as GC-MS, LC-MS/MS and HRMS, are required to develop multi-mycotoxin methods and to identify and quantify emerging, masked, and novel mycotoxins. Moreover, the high variety of samples involved in mycotoxin determination requires the development of appropriate extraction and clean-up techniques.

I hope that this Special Issue of Molecules will contribute to the advancement of analytical methods helpful in evaluating mycotoxin occurrence and diffusion in food and feed.

Prof. Dr. Terenzio Bertuzzi
Dr. Chiara Lanzanova
Dr. Sabrina Locatelli
Guest Editor

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Keywords

  • Mycotoxins
  • Analytical methods
  • Food safety
  • Monitoring

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Published Papers (7 papers)

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Research

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8 pages, 582 KiB  
Communication
Rapid Detection and Quantification of Patulin and Citrinin Contamination in Fruits
by Sudharsan Sadhasivam, Omer Barda, Varda Zakin, Ram Reifen and Edward Sionov
Molecules 2021, 26(15), 4545; https://doi.org/10.3390/molecules26154545 - 27 Jul 2021
Cited by 17 | Viewed by 3967
Abstract
Patulin (PAT) and citrinin (CTN) are the most common mycotoxins produced by Penicillium and Aspergillus species and are often associated with fruits and fruit by-products. Hence, simple and reliable methods for monitoring these toxins in foodstuffs are required for regular quality assessment. In [...] Read more.
Patulin (PAT) and citrinin (CTN) are the most common mycotoxins produced by Penicillium and Aspergillus species and are often associated with fruits and fruit by-products. Hence, simple and reliable methods for monitoring these toxins in foodstuffs are required for regular quality assessment. In this study, we aimed to establish a cost-effective method for detection and quantification of PAT and CTN in pome fruits, such as apples and pears, using high-performance liquid chromatography (HPLC) coupled with spectroscopic detectors without the need for any clean-up steps. The method showed good performance in the analysis of these mycotoxins in apple and pear fruit samples with recovery ranges of 55–97% for PAT and 84–101% for CTN, respectively. The limits of detection (LOD) of PAT and CTN in fruits were 0.006 µg/g and 0.001 µg/g, while their limits of quantification (LOQ) were 0.018 µg/g and 0.003 µg/g, respectively. The present findings indicate that the newly developed HPLC method provides rapid and accurate detection of PAT and CTN in fruits. Full article
(This article belongs to the Special Issue Analytical Methods for Mycotoxin Analysis)
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14 pages, 309 KiB  
Article
Incidence of Fusarium Mycotoxins in Wheat and Maize from Albania
by Dritan Topi, Janja Babič, Katarina Pavšič-Vrtač, Gabrijela Tavčar-Kalcher and Breda Jakovac-Strajn
Molecules 2021, 26(1), 172; https://doi.org/10.3390/molecules26010172 - 31 Dec 2020
Cited by 31 | Viewed by 3711
Abstract
In this study, ten Fusarium toxins were analysed in wheat and maize commodities from Albania. In total, 71 samples of wheat and 45 samples of maize were collected from different producing regions. The analytical procedure consisted of a simple one-step sample extraction followed [...] Read more.
In this study, ten Fusarium toxins were analysed in wheat and maize commodities from Albania. In total, 71 samples of wheat and 45 samples of maize were collected from different producing regions. The analytical procedure consisted of a simple one-step sample extraction followed by the determination of toxins using liquid chromatography coupled with tandem mass spectrometry. Fusarium toxins were found in 23% of the analysed wheat samples and in 78% of maize samples. In maize samples, most often fumonisins B1 (FB1) and B2 (FB2) were found. They were present in 76% of samples. They were detected in all positive samples except in one with concentrations ranging from 59.9 to 16,970 μg/kg. The sum of FB1 and FB2 exceeded the EU maximum permitted level (4000 μg/kg) in 31% of maize samples. In wheat samples, the only detected Fusarium mycotoxin was deoxynivalenol (DON), present in 23% of samples. In one sample with the concentration of 1916 μg/kg, the EU maximum permitted level (1250 μg/kg) was exceeded. This is the first report on the presence of Fusarium toxins in wheat and maize grains cultivated in Albania. Full article
(This article belongs to the Special Issue Analytical Methods for Mycotoxin Analysis)
18 pages, 3591 KiB  
Article
Evaluation of Two Fully Automated Setups for Mycotoxin Analysis Based on Online Extraction-Liquid Chromatography–Tandem Mass Spectrometry
by Edvaldo Vasconcelos Soares Maciel, Karen Mejía-Carmona and Fernando Mauro Lanças
Molecules 2020, 25(12), 2756; https://doi.org/10.3390/molecules25122756 - 15 Jun 2020
Cited by 19 | Viewed by 2905
Abstract
Mycotoxins are secondary metabolites of fungi species widely known for their potentially toxic effects on human health. Considering their frequent presence in crops and their processed food, monitoring them on food-based matrices is now an important topic. Within such a context, the sample [...] Read more.
Mycotoxins are secondary metabolites of fungi species widely known for their potentially toxic effects on human health. Considering their frequent presence in crops and their processed food, monitoring them on food-based matrices is now an important topic. Within such a context, the sample preparation step is usually mandatory before the chromatographic analysis, due to the complexity of matrices such as nuts, cereals, beverages, and others. For these reasons, we herein present the evaluation of two greener setups, based on the automation and miniaturization of the sample preparation step for mycotoxin analysis in different beverages. Firstly, we describe an analytical method based on a multidimensional assembly, coupling a lab-made microextraction column (508 µm i.d. × 100 mm) to a UPLC–MS/MS for the analysis of ochratoxin A in beverages. This configuration used a synthesized sorbent phase containing C18-functionalized graphene–silica particles, which exhibited excellent extraction performance, as well as being reusable and cheaper than commercially available extractive phases. Sequentially, a second setup, based on a multidimensional capillary LC coupled to MS/MS, was assessed for the same purpose. In this case, a graphene oxide-based capillary extraction column (254 µm i.d. × 200 mm) was used as the first dimension, while a C18 analytical capillary column performed the mycotoxin separation in beverages. Although this second one has similarities with the first, we focused mainly on the benefits related to the link between a miniaturized/automated sample preparation device with a capillary LC–MS/MS system, which made our analysis greener. Additionally, the chromatographic efficiency could even be enhanced. Full article
(This article belongs to the Special Issue Analytical Methods for Mycotoxin Analysis)
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13 pages, 1225 KiB  
Article
Co-Occurrence of Moniliformin and Regulated Fusarium Toxins in Maize and Wheat Grown in Italy
by Terenzio Bertuzzi, Paola Giorni, Silvia Rastelli, Patrizia Vaccino, Chiara Lanzanova and Sabrina Locatelli
Molecules 2020, 25(10), 2440; https://doi.org/10.3390/molecules25102440 - 23 May 2020
Cited by 16 | Viewed by 4265
Abstract
The co-occurrence of moniliformin (MON), fumonisins (FBs), and deoxynivalenol (DON) was evaluated in maize, durum, and common wheat grown in different experimental fields located in several Italian regions. MON was quantified using a LC-MS/MS method adding lanthanum ions in the mobile phase. In [...] Read more.
The co-occurrence of moniliformin (MON), fumonisins (FBs), and deoxynivalenol (DON) was evaluated in maize, durum, and common wheat grown in different experimental fields located in several Italian regions. MON was quantified using a LC-MS/MS method adding lanthanum ions in the mobile phase. In maize, MON contamination was widespread and considerable; the toxin was detected in almost all the samples (95.1%) and exceeded 500 and 1000 µg kg−1 in 42.0% and in 18.5% of samples, respectively. Significant positive correlation was found between MON and FB contamination levels. When there were not droughty climate conditions, a positive significant correlation was found between growing degree days (GDD) and MON values. In wheat, MON contamination was not widespread like in maize and it was lower in common wheat than in durum wheat. In durum wheat, MON was detected in 45.0% of the samples with only 6 samples (7.5%) exceeding 500 µg kg−1, while in common wheat the toxin was detected above the LOD in 18.7% of samples exceeding 100 µg kg−1 in only two samples (2.5%). No correlation was found with DON contamination. Climate conditions influenced both MON and DON occurrence. Full article
(This article belongs to the Special Issue Analytical Methods for Mycotoxin Analysis)
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11 pages, 1633 KiB  
Article
Identification and Distribution of Novel Metabolites of Lolitrem B in Mice by High-Resolution Mass Spectrometry
by Priyanka Reddy, Aaron Elkins, Joanne Hemsworth, Kathryn Guthridge, Simone Vassiliadis, Elizabeth Read, German Spangenberg and Simone Rochfort
Molecules 2020, 25(2), 372; https://doi.org/10.3390/molecules25020372 - 16 Jan 2020
Cited by 6 | Viewed by 3370
Abstract
Lolitrem B is the most potent indole-diterpene mycotoxin produced by Epichloë festucae var. lolii (termed LpTG-1), with severe intoxication cases reported in livestock. To date, there are no in vivo metabolism studies conducted for the mycotoxin. A mouse model assay established for [...] Read more.
Lolitrem B is the most potent indole-diterpene mycotoxin produced by Epichloë festucae var. lolii (termed LpTG-1), with severe intoxication cases reported in livestock. To date, there are no in vivo metabolism studies conducted for the mycotoxin. A mouse model assay established for assessing toxicity of indole-diterpenes was used to investigate metabolic products of lolitrem B. Mice were administered lolitrem B at 0.5 and 2.0 mg/kg body weight (b.wt) intraperitoneally before body and brain tissues were collected at 6 h and 24 h post-treatment. Samples were cryoground and subjected to a biphasic or monophasic extraction. The aqueous and lipophilic phases were analysed using liquid chromatography high-resolution mass spectrometry (LC–HRMS); data analysis was performed with Compound Discoverer™ software. A total of 10 novel phase I metabolic products were identified in the lipophilic phase and their distribution in the liver, kidney and various brain regions are described. The biotransformation products of lolitrem B were found to be present in low levels in the brain. Based on structure–activity postulations, six of these may contribute towards the protracted tremors exhibited by lolitrem B-exposed animals. Full article
(This article belongs to the Special Issue Analytical Methods for Mycotoxin Analysis)
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Review

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15 pages, 1230 KiB  
Review
T-2 Toxin—The Most Toxic Trichothecene Mycotoxin: Metabolism, Toxicity, and Decontamination Strategies
by Edyta Janik, Marcin Niemcewicz, Marcin Podogrocki, Michal Ceremuga, Maksymilian Stela and Michal Bijak
Molecules 2021, 26(22), 6868; https://doi.org/10.3390/molecules26226868 - 14 Nov 2021
Cited by 127 | Viewed by 10616
Abstract
Among trichothecenes, T-2 toxin is the most toxic fungal secondary metabolite produced by different Fusarium species. Moreover, T-2 is the most common cause of poisoning that results from the consumption of contaminated cereal-based food and feed reported among humans and animals. The food [...] Read more.
Among trichothecenes, T-2 toxin is the most toxic fungal secondary metabolite produced by different Fusarium species. Moreover, T-2 is the most common cause of poisoning that results from the consumption of contaminated cereal-based food and feed reported among humans and animals. The food and feed most contaminated with T-2 toxin is made from wheat, barley, rye, oats, and maize. After exposition or ingestion, T-2 is immediately absorbed from the alimentary tract or through the respiratory mucosal membranes and transported to the liver as a primary organ responsible for toxin's metabolism. Depending on the age, way of exposure, and dosage, intoxication manifests by vomiting, feed refusal, stomach necrosis, and skin irritation, which is rarely observed in case of mycotoxins intoxication. In order to eliminate T-2 toxin, various decontamination techniques have been found to mitigate the concentration of T-2 toxin in agricultural commodities. However, it is believed that 100% degradation of this toxin could be not possible. In this review, T-2 toxin toxicity, metabolism, and decontamination strategies are presented and discussed. Full article
(This article belongs to the Special Issue Analytical Methods for Mycotoxin Analysis)
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26 pages, 498 KiB  
Review
Reviewing the Analytical Methodologies to Determine the Occurrence of Citrinin and Its Major Metabolite, Dihydrocitrinone, in Human Biological Fluids
by Liliana Silva, André Pereira, Sofia Duarte, Angelina Pena and Celeste Lino
Molecules 2020, 25(12), 2906; https://doi.org/10.3390/molecules25122906 - 24 Jun 2020
Cited by 9 | Viewed by 2608
Abstract
Until now, the available data regarding citrinin (CIT) levels in food and the consumption of contaminated foods are insufficient to allow a reliable estimate of intake. Therefore, biomonitoring configuring analysis of parent compound and/or metabolites in biological fluids, such as urine or blood, [...] Read more.
Until now, the available data regarding citrinin (CIT) levels in food and the consumption of contaminated foods are insufficient to allow a reliable estimate of intake. Therefore, biomonitoring configuring analysis of parent compound and/or metabolites in biological fluids, such as urine or blood, is being increasingly applied in the assessment of human exposure to CIT and its metabolite, dihydrocitrinone (DH-CIT). Most studies report urinary levels lower for the parent compound when compared with DH-CIT. A high variability either in the mean levels or in the inter-individual ratios of CIT/DH-CIT between the reported studies has been found. Levels of DH-CIT in urine were reported as being comprised between three to seventeen times higher than the parent mycotoxin. In order to comply with this objective, sensitive analytical methodologies for determining biomarkers of exposure are required. Recent development of powerful analytical techniques, namely liquid chromatography coupled to mass spectrometry (LC-MS/MS) and ultra-high-performance liquid chromatography (UHPLC-MS/MS) have facilitated biomonitoring studies, mainly in urine samples. In the present work, evidence on human exposure to CIT through its occurrence and its metabolite, in biological fluids, urine and blood/plasma, in different countries, is reviewed. The analytical methodologies usually employed to evaluate trace quantities of these two molecules, are also presented. In this sense, relevant data on sampling (size and pre-treatment), extraction, cleanup and detection and quantification techniques and respective chromatographic conditions, as well as the analytical performance, are evidenced. Full article
(This article belongs to the Special Issue Analytical Methods for Mycotoxin Analysis)
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