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Toxins
Special Issues
Multi Methods for Detecting Natural Toxins
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Multi Methods for Detecting Natural Toxins

A special issue of Toxins (ISSN 2072-6651).

Deadline for manuscript submissions: 30 April 2025 | Viewed by 13373

Special Issue Editor

Dr. Svetlana V. Malysheva

E-Mail Website
Guest Editor
Unit Toxins, Service Organic Contaminants and Additives, Scientific Directorate Chemical and Physical Health Risks, Leuvensesteenweg 17, 3080 Tervuren, Belgium
Interests: analytical chemistry; method development and validation; mass spectrometry; plant toxins; food and feed analysis; food safety

Special Issue Information

Dear Colleagues,

Natural toxins are a diverse group of toxic compounds that are naturally produced by living organisms. Possessing various structures, natural toxins demonstrate different biological functions and toxicity. While these compounds are not harmful to the organisms producing them, via different exposure routes, they can reach human and animal bodies and can cause adverse health effects. Exposure to natural toxins needs to be studied and monitored; therefore, the availability of robust, sensitive and fit-for-purpose analytical methods for the detection, screening and/or quantification of various groups of natural toxins is crucial.

This Special Issue is dedicated to the analysis of natural toxins via multi-analyte methods. The simultaneous detection of multiple toxins belonging to the same class, as well as those belonging to different classes (or origins) of toxins, is emphasized. The applicative fields are diverse and can be related to food and feed control, water quality, environmental contamination, biomonitoring and other applications involving multi-toxin analytical methods. It is vital that the methods presented can demonstrate improvements in or the novelty of the analytical approach (e.g., analytes of interest, detection technique, sample preparation, application, performance…).

Dr. Svetlana V. Malysheva
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a double-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Toxins is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • fungal toxins
  • plant toxins
  • bacterial toxins
  • marine and freshwater toxins
  • method development
  • analysis
  • multi-toxin methods
  • validation

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Published Papers (7 papers)

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Research

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15 pages, 1038 KiB  
Article
Determination of 16 Hydroxyanthracene Derivatives in Food Supplements Using LC-MS/MS: Method Development and Application
by Svetlana V. Malysheva, Benoît Guillaume, Céline Vanhee and Julien Masquelier
Toxins 2024, 16(12), 505; https://doi.org/10.3390/toxins16120505 - 23 Nov 2024
Viewed by 886
Abstract
Hydroxyanthracene derivatives (HADs) are plant substances produced by a variety of plant species, including different Aloe, Rheum, and Rhamnus species and Cassia senna. These plants are often used in food supplements to improve bowel function. However, recently, the European Commission prohibited [...] Read more.
Hydroxyanthracene derivatives (HADs) are plant substances produced by a variety of plant species, including different Aloe, Rheum, and Rhamnus species and Cassia senna. These plants are often used in food supplements to improve bowel function. However, recently, the European Commission prohibited a number of HADs due to toxicological concerns. These HADs included aloin (aloin A and aloin B), aloe-emodin, emodin, and danthron. Most of the currently available analytical methods are restricted to the analysis of only these compounds and do not include other HADs. In this view, a multi-analyte method could be useful for both regulatory analysis and dietary intake studies. To this end, such a method, employing liquid chromatography–tandem mass spectrometry and targeting 16 different HADs, was developed and validated in this study. Limits of quantification were in the range from 0.025 mg kg−1 to 1 mg kg−1. The recovery of the method was within the acceptable range of 80% to 120%, with the exception of physcion. Repeatability varied from 0.5% to 11.6%, and the range for within-laboratory reproducibility was from 3.4% to 16.3%. The expanded measurement uncertainty was below 50% for all HADs. Subsequently, 24 commercial samples of food supplements and herbal infusions sourced in Belgium were analyzed. The results indicated that although the industry put a great effort into minimizing the amount of aloin and danthron present in food supplements, more than half of the products still exceeded the maximum tolerated levels suggested for aloe-emodin and emodin. Full article
(This article belongs to the Special Issue Multi Methods for Detecting Natural Toxins)
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15 pages, 646 KiB  
Article
Validation of a UPLC-MS/MS Method for Multi-Matrix Biomonitoring of Alternaria Toxins in Humans
by Lia Visintin, María García Nicolás, Sarah De Saeger and Marthe De Boevre
Toxins 2024, 16(7), 296; https://doi.org/10.3390/toxins16070296 - 28 Jun 2024
Viewed by 992
Abstract
Mycotoxins, natural toxins produced by fungi, contaminate nearly 80% of global food crops. Alternaria mycotoxins, including alternariol (AOH), alternariol monomethylether (AME), and tenuazonic acid (TeA), present a health concern due to their prevalence in various plants and fruits. Exposure to these toxins exceeds [...] Read more.
Mycotoxins, natural toxins produced by fungi, contaminate nearly 80% of global food crops. Alternaria mycotoxins, including alternariol (AOH), alternariol monomethylether (AME), and tenuazonic acid (TeA), present a health concern due to their prevalence in various plants and fruits. Exposure to these toxins exceeds the threshold of toxicological concern in some European populations, especially infants and toddlers. Despite this, regulatory standards for Alternaria toxins remain absent. The lack of toxicokinetic parameters, reference levels, and sensitive detection methods complicates risk assessment and highlights the necessity for advanced biomonitoring (HBM) techniques. This study addresses these challenges by developing and validating ultra-high performance liquid chromatography method coupled with tandem mass spectrometry to quantify AOH, AME, TeA, and their conjugates in multiple biological matrices. The validated method demonstrates robust linearity, precision, recovery (94–111%), and sensitivity across urine (LOD < 0.053 ng/mL), capillary blood (LOD < 0.029 ng/mL), and feces (LOD < 0.424 ng/g), with significantly lower LOD for TeA compared to existing methodologies. The application of minimally invasive microsampling techniques for the blood collection enhances the potential for large-scale HBM studies. These advancements represent a step toward comprehensive HBM and exposure risk assessments for Alternaria toxins, facilitating the generation of data for regulatory authorities. Full article
(This article belongs to the Special Issue Multi Methods for Detecting Natural Toxins)
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13 pages, 13323 KiB  
Article
Sensitive Detection and Differentiation of Biologically Active Ricin and Abrin in Complex Matrices via Specific Neutralizing Antibody-Based Cytotoxicity Assay
by Zhi Li, Bo Ma, Mengqiang Gong, Lei Guo, Lili Wang, Hua Xu and Jianwei Xie
Toxins 2024, 16(6), 237; https://doi.org/10.3390/toxins16060237 - 23 May 2024
Viewed by 1165
Abstract
Ricin and abrin are highly potent plant-derived toxins, categorized as type II ribosome-inactivating proteins. High toxicity, accessibility, and the lack of effective countermeasures make them potential agents in bioterrorism and biowarfare, posing significant threats to public safety. Despite the existence of many effective [...] Read more.
Ricin and abrin are highly potent plant-derived toxins, categorized as type II ribosome-inactivating proteins. High toxicity, accessibility, and the lack of effective countermeasures make them potential agents in bioterrorism and biowarfare, posing significant threats to public safety. Despite the existence of many effective analytical strategies for detecting these two lethal toxins, current methods are often hindered by limitations such as insufficient sensitivity, complex sample preparation, and most importantly, the inability to distinguish between biologically active and inactive toxin. In this study, a cytotoxicity assay was developed to detect active ricin and abrin based on their potent cell-killing capability. Among nine human cell lines derived from various organs, HeLa cells exhibited exceptional sensitivity, with limits of detection reaching 0.3 ng/mL and 0.03 ng/mL for ricin and abrin, respectively. Subsequently, toxin-specific neutralizing monoclonal antibodies MIL50 and 10D8 were used to facilitate the precise identification and differentiation of ricin and abrin. The method provides straightforward and sensitive detection in complex matrices including milk, plasma, coffee, orange juice, and tea via a simple serial-dilution procedure without any complex purification and enrichment steps. Furthermore, this assay was successfully applied in the unambiguous identification of active ricin and abrin in samples from OPCW biotoxin exercises. Full article
(This article belongs to the Special Issue Multi Methods for Detecting Natural Toxins)
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18 pages, 2078 KiB  
Article
New Analytical Approach to Quinolizidine Alkaloids and Their Assumed Biosynthesis Pathways in Lupin Seeds
by Dvory Namdar, Patrick P. J. Mulder, Eyal Ben-Simchon, Yael Hacham, Loai Basheer, Ofer Cohen, Marcelo Sternberg and Oren Shelef
Toxins 2024, 16(3), 163; https://doi.org/10.3390/toxins16030163 - 21 Mar 2024
Cited by 4 | Viewed by 2363
Abstract
Alkaloids play an essential role in protecting plants against herbivores. Humans can also benefit from the pharmacological effects of these compounds. Plants produce an immense variety of structurally different alkaloids, including quinolizidine alkaloids, a group of bi-, tri-, and tetracyclic compounds produced by [...] Read more.
Alkaloids play an essential role in protecting plants against herbivores. Humans can also benefit from the pharmacological effects of these compounds. Plants produce an immense variety of structurally different alkaloids, including quinolizidine alkaloids, a group of bi-, tri-, and tetracyclic compounds produced by Lupinus species. Various lupin species produce different alkaloid profiles. To study the composition of quinolizidine alkaloids in lupin seeds, we collected 31 populations of two wild species native to Israel, L. pilosus and L. palaestinus, and analyzed their quinolizidine alkaloid contents. Our goal was to study the alkaloid profiles of these two wild species to better understand the challenges and prospective uses of wild lupins. We compared their profiles with those of other commercial and wild lupin species. To this end, a straightforward method for extracting alkaloids from seeds and determining the quinolizidine alkaloid profile by LC–MS/MS was developed and validated in-house. For the quantification of quinolizidine alkaloids, 15 analytical reference standards were used. We used GC–MS to verify and cross-reference the identity of certain alkaloids for which no analytical standards were available. The results enabled further exploration of quinolizidine alkaloid biosynthesis. We reviewed and re-analyzed the suggested quinolizidine alkaloid biosynthesis pathway, including the relationship between the amino acid precursor l-lysine and the different quinolizidine alkaloids occurring in seeds of lupin species. Revealing alkaloid compositions and highlighting some aspects of their formation pathway are important steps in evaluating the use of wild lupins as a novel legume crop. Full article
(This article belongs to the Special Issue Multi Methods for Detecting Natural Toxins)
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10 pages, 1362 KiB  
Article
Magnetic Molecularly Imprinted Polymers for Selective Extraction of Aflatoxins from Feeds
by María del Carmen Pérez-Álvarez, Natalia Arroyo-Manzanares, Natalia Campillo and Pilar Viñas
Toxins 2024, 16(3), 120; https://doi.org/10.3390/toxins16030120 - 29 Feb 2024
Viewed by 1690
Abstract
Magnetic molecularly imprinted polymers (MMIPs) have fused molecular imprinting technology with magnetic separation technology, emerging as an innovative material capable of recognizing specific molecules and efficiently separating target substances. Their application to the extraction and purification of mycotoxins has great potential, due to [...] Read more.
Magnetic molecularly imprinted polymers (MMIPs) have fused molecular imprinting technology with magnetic separation technology, emerging as an innovative material capable of recognizing specific molecules and efficiently separating target substances. Their application to the extraction and purification of mycotoxins has great potential, due to the toxicity and economic impact of these contaminants. In this work, MMIP has been proposed as a sample treatment for the determination of main four aflatoxins (B1, B2, G1 and G2) in pig feed. The MMIP was formed through the integration of magnetic material (Fe3O4) with commercial molecularly imprinted polymers, avoiding the synthesis step and, therefore, simplifying the process. The analyses were carried out by high-performance liquid chromatography with fluorescence detection and the method was validated and limits of quantification (LOQs) between 0.09 and 0.47 ng/g were obtained, below the allowed or recommended levels by the European Union. Repeatability and intermediate precision showed relative standard deviations lower than 10% in all cases and trueness ranged from 92 to 111%. Finally, the proposed method was applied to 31 real pig feed samples, detecting aflatoxins with concentrations between 0.2 and 3.2 ng/g. Full article
(This article belongs to the Special Issue Multi Methods for Detecting Natural Toxins)
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13 pages, 855 KiB  
Article
Validation of a Targeted LC–MS/MS Method for Cereulide and Application in Food and Faeces
by Julien Masquelier, Céline Segers, Bram Jacobs, Tom Van Nieuwenhuysen, Laurence Delbrassinne and Els Van Hoeck
Toxins 2024, 16(1), 13; https://doi.org/10.3390/toxins16010013 - 27 Dec 2023
Cited by 1 | Viewed by 2088
Abstract
Cereulide is an emetic toxin produced by some strains of Bacillus cereus. This bacterial toxin, a cyclic 1.2 kDa dodecadepsipeptide, is stable to heat and acids and causes nausea and vomiting when ingested via contaminated food. This work aimed to develop and [...] Read more.
Cereulide is an emetic toxin produced by some strains of Bacillus cereus. This bacterial toxin, a cyclic 1.2 kDa dodecadepsipeptide, is stable to heat and acids and causes nausea and vomiting when ingested via contaminated food. This work aimed to develop and validate a targeted analytical method applying liquid chromatography-tandem mass spectrometry (LC–MS/MS) to quantify this toxin in food and human faeces. Samples were extracted with acetonitrile in the presence of 13C6-cereulide, a labelled internal standard, and purified by centrifugation and filtration. The limits of quantification were 0.5 and 0.3 µg kg−1 for food and faeces, respectively. The linearity of the method was very good, with calculated R2 values above 0.995. The mean recovery of the method was within the acceptable range of 70.0%–120.0%, the repeatability was not higher than 7.3%, and the highest intra-laboratory reproducibility was 8.9%. The estimated range for the expanded measurement uncertainty was between 5.1% and 18.0%. The LC–MS/MS method was used to analyse one food sample (rice) from a Belgian foodborne outbreak and five faecal samples from patients with clinical symptoms after consumption of the contaminated rice. The levels of cereulide were 12.22 µg g−1 for food and between 6.32 and 773.37 ng g−1 for faecal samples. Full article
(This article belongs to the Special Issue Multi Methods for Detecting Natural Toxins)
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Review

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18 pages, 633 KiB  
Review
Challenges of Diphtheria Toxin Detection
by Marta Prygiel, Ewa Mosiej, Maciej Polak, Katarzyna Krysztopa-Grzybowska, Karol Wdowiak, Kamila Formińska and Aleksandra A. Zasada
Toxins 2024, 16(6), 245; https://doi.org/10.3390/toxins16060245 - 26 May 2024
Viewed by 2809
Abstract
Diphtheria toxin (DT) is the main virulence factor of Corynebacterium diphtheriae, C. ulcerans and C. pseudotuberculosis. Moreover, new Corynebacterium species with the potential to produce diphtheria toxin have also been described. Therefore, the detection of the toxin is the most important test in [...] Read more.
Diphtheria toxin (DT) is the main virulence factor of Corynebacterium diphtheriae, C. ulcerans and C. pseudotuberculosis. Moreover, new Corynebacterium species with the potential to produce diphtheria toxin have also been described. Therefore, the detection of the toxin is the most important test in the microbiological diagnosis of diphtheria and other corynebacteria infections. Since the first demonstration in 1888 that DT is a major virulence factor of C. diphtheriae, responsible for the systemic manifestation of the disease, various methods for DT detection have been developed, but the diagnostic usefulness of most of them has not been confirmed on a sufficiently large group of samples. Despite substantial progress in the science and diagnostics of infectious diseases, the Elek test is still the basic recommended diagnostic test for DT detection. The challenge here is the poor availability of an antitoxin and declining experience even in reference laboratories due to the low prevalence of diphtheria in developed countries. However, recent and very promising assays have been developed with the potential for use as rapid point-of-care testing (POCT), such as ICS and LFIA for toxin detection, LAMP for tox gene detection, and biosensors for both. Full article
(This article belongs to the Special Issue Multi Methods for Detecting Natural Toxins)
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