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Daily dietary intake inevitably exposes individuals to various natural toxins, which may pose potential health threats. Focusing only on specific toxins could underestimate dietary risks. Therefore, we have developed a suspect and nontarget method based on ultrahigh-performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) to screen both known and unknown natural toxins in various foodstuffs. An in-house database containing 2952 natural toxins including fungal toxins, phytotoxins, animal toxins and cyanotoxins was established, facilitating suspect screening. Predicted retention time and mass spectrometry data were employed to enhance the confidence levels. Subsequently, Nontarget screening method was conducted based on molecular network analysis, annotating the edges and nodes through modified types and fragmentation characteristics. Finally, we analyzed 102 foodstuff samples and identified a total of 90 natural toxins, including mycotoxins and phytotoxins, with 65 identified by suspect screening and 25 by nontarget screening. Based on measured concentrations, the daily per capita dietary intake of total natural toxins was estimated, it was below risk doses for natural toxins with available reference values. Overall, this work established a novel method for the comprehensive identification of natural toxins in foodstuffs and emphasized the importance of dietary risk assessment for natural toxins., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Botulinum neurotoxins (BoNTs), ricin, and many other biological toxins are called AB toxins possessing heterogeneous A and B subunits. We propose herein a quick and safe sensing approach to AB toxins based on their unique quaternary structures. The proposed approach utilizes IgG antibodies against their A-subunits in combination with those human cell-membrane glycolipids that act as the natural ligands of B-subunits. In practice, an IgG antibody against the A-subunit of a target toxin is selected from commercially available sources and immobilized on the surface of Au nanoparticles to constitute a multivalent IgG/Au nanoconjugate. The derived IgG/Au conjugate is used in the pretreatment process of test samples for deactivating biological toxins in the form of a ternary toxin/antibody/Au complex. This process is implemented in advance to reduce the risk of handling biological toxins in laboratory work. On the other hand, the human glycolipid is immobilized on a tiny glass plate and used as a biosensor chip. The biosensor chip is set in the chamber of a flow sensing system using localized surface plasmon resonance (LSPR) spectrometry available in portable size at relatively low cost. In principle, the LSPR sensing system enables us to perform a rapid and selective detection for different kinds of biological toxins if the human glycolipid is correctly selected and installed in the sensing system. In the present LSPR sensing approach, a target AB toxin may have been deactivated during the pretreatment process. The test sample containing the deactivated AB toxin becomes a real target to be analyzed by the sensing system. In the present, we describe the concept of employing the commercially available IgG antibody in the pretreatment process followed by a typical procedure for converting it into the multivalent antibody/Au nanoconjugate and its preliminary applications in the LSPR detection of a ricin homologue (RCA 120 ) and BoNTs in different serotypes. The tested LSPR sensing approach has worked very well for the ricin homologue and certain serotypes of botulinum neurotoxins like BoNT/A, indicating that the prior deactivation process at their A-domains causes no significant damage to the function of their B-domains with respect to determining the host cell-membrane glycolipid. The experimental results also indicated that LSPR responses from these pretreated AB toxins are significantly amplified. That is obviously thanks to the presence of Au nanoparticles in the multivalent IgG/Au nanoconjugate. We suggest in conclusion that the proposed LSPR sensing approach will provide us with a safe and useful tool for the study of biological AB toxins based on their unique quaternary protein structures.

Rapid detection of uremic toxins is crucial due to their severe health risks, including oxidative stress, inflammation, neurotoxicity, cardiovascular complications, and progression of chronic kidney disease. Surface-enhanced Raman spectroscopy (SERS) may provide sensitive, fast, and clinical-grade real-time monitoring of these toxins, enabling effective management with timely dialysis treatments. This study introduces a 3D-printed microchamber that integrates the fabrication of plasmonic metal nanoparticles for the in-flow detection of biological toxins and pharmaceutical drugs using SERS, making it ideal for on-site diagnostics in clinical settings. The microchamber supports quantitative and highly reproducible detection with liquid volumes under 100 μL, which is crucial for trace-level biomarker detection and minimizing cross-contamination. It employs a tunable solvent exchange method for the in situ synthesis of silver nanoparticles (AgNPs) on flexible PDMS or rigid Si wafer substrates, avoiding costly nanofabrication techniques. Ultralow detection limits were achieved for two model compounds and three pharmaceutical drugs: 10 -11 M for rhodamine 6G, 10 -7 M for adenine, and 10 -6 M for the pharmaceutical drugs. A total of 13 biological toxins, including three neurotransmitters, one neuromodulator, five amino acids, two polyamines, and two urea cycle metabolites, were detected with quantitative limits ranging from 10 -3 to 10 -6 M, all below permissible levels and aligning with physiological conditions. SERS detection within microchambers facilitates rapid on-site analysis, proving ideal for personalized health monitoring, point-of-care diagnostics, and environmental pollution assessment.

Ambient ionisation mass spectrometry (AIMS) is a form of mass spectrometry whereby analyte ionisation occurs outside of a vacuum source under ambient conditions. This enables the direct analysis of samples in their native state, with little or no sample preparation and without chromatographic separation. The removal of these steps facilitates a much faster analytical process, enabling the direct analysis of samples within minutes if not seconds. Consequently, AIMS has gained rapid popularity across a diverse range of applications, in particular the analysis of drugs and toxins. Numerous fields rely upon mass spectrometry for the detection and identification of drugs, including clinical diagnostics, forensic chemistry, and food safety. However, all of these fields are hindered by the time-consuming and laboratory-confined nature of traditional techniques. As such, the potential for AIMS to resolve these challenges has resulted in a growing interest in ambient ionisation for drug and toxin analysis. Since the early 2000s, forensic science, diagnostic testing, anti-doping, pharmaceuticals, environmental analysis and food safety have all seen a marked increase in AIMS applications, foreshadowing a new future for drug testing. In this review, some of the most promising AIMS techniques for drug analysis will be discussed, alongside different applications of AIMS published over a 5-year period, to provide a summary of the recent research activity for ambient ionisation for drug and toxin analysis., (© 2024 The Authors. Drug Testing and Analysis published by John Wiley & Sons Ltd.)

Biotoxins are pervasive in food and the environment, posing significant risk to human health. The most effective strategy to mitigate the risk arising from biotoxin exposure is through their specific and sensitive detection. Aptasensors have emerged as pivotal tools, leveraging aptamers as biorecognition elements to transduce the specificity of aptamer-target interactions into quantifiable signals for analytical applications, thereby facilitating the meticulous detection of biotoxins. When integrated with readily portable devices such as lateral flow assays (LFAs), personal glucose meters (PGMs), smartphones, and various meters measuring parameters like pH and pressure, aptasensors have significantly advanced the field of biotoxin monitoring. These commercially available devices enable precise, in situ, and real-time analysis, offering great potential for portable biotoxin detection in food and environmental matrices. This review highlights the recent progress in biotoxin monitoring using portable aptasensors, discussing both their potential applications and the challenges encountered. By addressing these impediments, we anticipate that a portable aptasensor-based detection system will open new avenues in biotoxin monitoring in the future.

Biological weapons, primarily dispersed as aerosols, can spread not only to the targeted area but also to adjacent regions following the movement of air driven by wind. Thus, there is a growing demand for toxin analysis because biological weapons are among the most influential and destructive. Specifically, such a technique should be hand-held, rapid, and easy to use because current methods require more time and well-trained personnel. Our study demonstrates the use of a novel lateral flow immunoassay, which has a confined structure like a double barbell in the detection area (so called c-LFA) for toxin detection such as staphylococcal enterotoxin B (SEB), ricinus communis (Ricin), and botulinum neurotoxin type A (BoNT-A). Additionally, we have explored the integration of machine learning (ML), specifically, a toxin chip boosting (TOCBoost) hybrid algorithm for improved sensitivity and specificity. Consequently, the ML powered c-LFA concurrently categorized three biological toxin types with an average accuracy as high as 95.5%. To our knowledge, the sensor proposed in this study is the first attempt to utilize ML for the assessment of toxins. The advent of the c-LFA orchestrated a paradigm shift by furnishing a versatile and robust platform for the rapid, on-site detection of various toxins, including SEB, Ricin, and BoNT-A. Our platform enables accessible and on-site toxin monitoring for non-experts and can potentially be applied to biosecurity.

Pyrrolizidine alkaloids (PAs) are toxic compounds that occur naturally in certain plants, however, there are many secondary pathways causing PA contamination of other plants, including medicinal herbs and plant-based food products, which pose a risk of human intoxication. It is proven that chronic exposure to PAs causes serious adverse health consequences resulting from their cytotoxicity and genotoxicity. This review briefly presents PA occurrence, structures, chemistry, and toxicity, as well as a set of analytical methods. Recently developed sensitive electrochemical and chromatographic methods for the determination of PAs in honey, teas, herbs, and spices were summarized. The main strategies for improving the analytical efficiency of PA determination are related to the use of mass spectrometric (MS) detection; therefore, this review focuses on advances in MS-based methods. Raising awareness of the potential health risks associated with the presence of PAs in food and herbal medicines requires ongoing research in this area, including the development of sensitive methods for PA determination and rigorous legal regulations of PA intake from herbal products. The maximum levels of PAs in certain products are regulated by the European Commission; however, the precise knowledge about which products contain trace but significant amounts of these alkaloids is still insufficient.

Consumers in developed and Western European countries are becoming more aware of the impact of food on their health, and they demand clear, transparent, and reliable information from the food industry about the products they consume. They recognise that food safety risks are often due to the unexpected presence of contaminants throughout the food supply chain. Among these, mycotoxins produced by food-infecting fungi, endogenous toxins from certain plants and organisms, pesticides, and other drugs used excessively during farming and food production, which lead to their contamination and accumulation in foodstuffs, are the main causes of concern. In this context, the goals of this review are to provide a comprehensive overview of the presence of toxic molecules reported in foodstuffs since 2020 through the Rapid Alert System for Food and Feed (RASFF) portal and use chromatography to address this challenge. Overall, natural toxins, environmental pollutants, and food-processing contaminants are the most frequently reported toxic molecules, and liquid chromatography and gas chromatography are the most reliable approaches for their control. However, faster, simpler, and more powerful analytical procedures are necessary to cope with the growing pressures on the food chain supply.

Naja nivea ( N. nivea ) is classed as a category one snake by the World Health Organization since its envenomation causes high levels of mortality and disability annually. Despite this, there has been little research into the venom composition of N. nivea , with only one full venom proteome published to date. Our current study separated N. nivea venom using size exclusion chromatography before utilizing a traditional bottom-up proteomics approach to unravel the composition of the venom proteome. As expected by its clinical presentation, N. nivea venom was found to consist mainly of neurotoxins, with three-finger toxins (3FTx), making up 76.01% of the total venom proteome. Additionally, cysteine-rich secretory proteins (CRISPs), vespryns (VESPs), cobra venom factors (CVFs), 5'-nucleotidases (5'NUCs), nerve growth factors (NGFs), phospholipase A2s (PLA 2 ), acetylcholinesterases (AChEs), Kunitz-type serine protease inhibitor (KUN), phosphodiesterases (PDEs), L-amino acid oxidases (LAAOs), hydrolases (HYDs), snake venom metalloproteinases (SVMPs), and snake venom serine protease (SVSP) toxins were also identified in decreasing order of abundance. Interestingly, contrary to previous reports, we find PLA 2 toxins in N. nivea venom. This highlights the importance of repeatedly profiling the venom of the same species to account for intra-species variation. Additionally, we report the first evidence of covalent protein complexes in N. nivea venom, which likely contribute to the potency of this venom.

In food safety analysis, the detection and control of foodborne pathogens and their toxins are of great importance. Monitoring of virus transmission is equally important, especially in light of recent findings that coronaviruses have been detected in frozen foods and packages during the current global epidemic of coronavirus disease 2019. In recent years, field-effect transistor (FET) biosensors have attracted considerable scholarly attention for pathogenic microorganisms and toxins detection and sensing due to their rapid response time, high sensitivity, wide dynamic range, high specificity, label-free detection, portability, and cost-effectiveness. FET-based biosensors can be modified with specific recognition elements, thus providing real-time qualitative and semiquantitative analysis. Furthermore, with advances in nanotechnology and device design, various high-performance nanomaterials are gradually applied in the detection of FET-based biosensors. In this article, we review specific detection in different biological recognition elements are immobilized on FET biosensors for the detection of pathogenic microorganisms and toxins, and we also discuss nonspecific detection by FET biosensors. In addition, there are still unresolved challenges in the development and application of FET biosensors for achieving efficient, multiplexed, in situ detection of pathogenic microorganisms and toxins. Therefore, directions for future FET biosensor research and applications are discussed.

Natural toxins (NTs) are poisonous secondary metabolites produced by living organisms developed to ward off predators. Especially low molecular weight NTs (MW<∼1 kDa), such as mycotoxins, phycotoxins, and plant toxins, are considered an important and growing food safety concern. Therefore, accurate risk assessment of food and feed for the presence of NTs is crucial. Currently, the analysis of NTs is predominantly performed with targeted high pressure liquid chromatography tandem mass spectrometry (HPLC-MS/MS) methods. Although these methods are highly sensitive and accurate, they are relatively expensive and time-consuming, while unknown or unexpected NTs will be missed. To overcome this, novel on-site screening methods and non-targeted HPLC high resolution mass spectrometry (HRMS) methods have been developed. On-site screening methods can give non-specialists the possibility for broad "scanning" of potential geographical regions of interest, while also providing sensitive and specific analysis at the point-of-need. Non-targeted chromatography-HRMS methods can detect unexpected as well as unknown NTs and their metabolites in a lab-based approach. The aim of this chapter is to provide an insight in the recent advances, challenges, and perspectives in the field of NTs analysis both from the on-site and the laboratory perspective., (Copyright © 2024. Published by Elsevier Inc.)

European vipers (genus Vipera ) are medically important snakes displaying considerable venom variation, occurring at different levels in this group. The presence of intraspecific venom variation, however, remains understudied in several Vipera species. Vipera seoanei is a venomous snake endemic to the northern Iberian Peninsula and south-western France, presenting notable phenotypic variation and inhabiting several diverse habitats across its range. We analysed the venoms of 49 adult specimens of V. seoanei from 20 localities across the species' Iberian distribution. We used a pool of all individual venoms to generate a V. seoanei venom reference proteome, produced SDS-PAGE profiles of all venom samples, and visualised patterns of variation using NMDS. By applying linear regression, we then assessed presence and nature of venom variation between localities, and investigated the effect of 14 predictors (biological, eco-geographic, genetic) on its occurrence. The venom comprised at least 12 different toxin families, of which five (i.e., PLA 2 , svSP, DI, snaclec, svMP) accounted for about 75% of the whole proteome. The comparative analyses of the SDS-PAGE venom profiles showed them to be remarkably similar across the sampled localities, suggesting low geographic variability. The regression analyses suggested significant effects of biological and habitat predictors on the little variation we detected across the analysed V. seoanei venoms. Other factors were also significantly associated with the presence/absence of individual bands in the SDS-PAGE profiles. The low levels of venom variability we detected within V. seoanei might be the result of a recent population expansion, or of processes other than directional positive selection.

Background: Various food commodities are vulnerable to different types of fungal pathogens and could be contaminated with differential classes of mycotoxins as a result. It is ideal to implement a generic method for the simultaneous determination of multi-mycotoxins in different food matrixes or agricultural products., Objective: In this study, a simplified sample preparation procedure and a reliable LC-MS/MS analytical method were developed for the comprehensive measurement of 37 regulated and emerging mycotoxins including five Alternaria toxins (ATs) and six major ergot alkaloids (EAs) and their corresponding epimers. Four different food commodities (baby wheat cereal, peanut, tomato puree, and blended flour) were chosen for method validation to demonstrate the applicability of this analytical method across a wide range of food types., Methods: Sample extraction was performed using a formic acid-acidified acetonitrile-water (4 + 1, v/v) solution followed by extract dry-down and reconstitution in a water-methanol (1 + 1, v/v) solution for analysis on a biphenyl LC column. Chromatographic analysis was performed using regular acidic LC conditions for baseline separation of ergot alkaloid epimers and completed with a short 11 min cycle time., Results: Accurate quantification was achieved using matrix-matched calibration standards in the range of 0.4 to 400 µg/kg. The recoveries of all mycotoxins (except citrinin) in fortified samples were from 70 to 120%, and the RSD was less than 20%., Conclusion: The established workflow was simple and fast for multi-mycotoxin determination in a wide variety of food commodities with LOQs needed to meet the regulatory levels., Highlights: The developed method provided the unique benefit of simultaneous analysis of Alternaria toxins (ATs) and ergot alkaloids (EAs) together with other major regulated mycotoxins., (© The Author(s) 2022. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

In paralytic shellfish toxin-producing dinoflagellates, intracellular levels of saxitoxin and its analogues (STXs) are controlled by a balance between degradation and biosynthesis in response to marine environmental fluctuations and stresses. The purpose of this study was to demonstrate the utility of statistical analysis of in vivo labeling data for the dynamic analysis of variations in toxin production under stress. A toxic strain of the dinoflagellate Alexandrium pacificum (Group IV) was cultured in colchicine-containing 15 N-labeled sodium nitrate-medium and metabolite levels were analyzed over time by liquid chromatography-mass spectrometry. Quantitative values of all isotopomers of precursor amino acids, biosynthetic intermediates, and major STXs were subjected to statistical analysis. The decrease of the nitrogen incorporation rates for all compounds suggested that colchicine decreased nitrate assimilation upstream of glutamate biosynthesis. In colchicine-treated cultures, the per-cell content of total STX analogues did not change significantly over time; however, the production rate of each pathway varied greatly. De novo STX biosynthesis was decreased by colchicine until Day 3, while the salvage pathway was not. Subsequently, biosynthesis by both pathways was enhanced. This analysis of dynamic metabolism provides new insights into the complex mechanisms regulating STX metabolism in dinoflagellates., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

As a filamentous and spoilage fungus, Alternaria spp. can not only infect processing tomatoes, but also produce a variety of mycotoxins which harm the health of human beings. To explore the production of Alternaria toxins in processing tomatoes during growth and storage, four main Alternaria toxins and four conjugated toxins were detected by ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and ultra-performance liquid chromatography-ion mobility quadrupole time-of-flight mass spectrometry (UPLC-IMS QToF MS) in processing tomatoes on different days after being inoculated with A. alternata . The results show that the content of Alternaria toxins in an in vivo assay is higher than that under field conditions. Tenuazonic acid (TeA) is the predominant toxin detected in the field (205.86~41,389.19 μg/kg) and in vivo (7.64~526,986.37 μg/kg) experiments, and the second-most abundant toxin is alternariol (AOH). In addition, a small quantity of conjugated toxins, AOH-9-glucoside (AOH-9-Glc) and alternariol monomethyl ether-3-glucoside (AME-3-Glc), were screened in the in vivo experiment. This is the first time the potential of Alternaria toxins produced in tomatoes during the harvest period has been studied in order to provide data for the prevention and control of Alternaria toxins.

Recently, demand for fermented foods has increased due to their improved nutritional value, taste, and health-promoting properties. Worldwide consumption of these products is increasing. Fermented foods are generally safe for human consumption. However, some toxins, primarily biogenic amines (putrescine, phenylethylamine, histamine, tyramine, and cadaverine), mycotoxins (fumonisins, aflatoxins, ochratoxin A, zearalenone, and trichothecenes), and bacterial toxins (endotoxins, enterotoxins, and emetic toxins) can be produced as a result of using an inappropriate starter culture, processing conditions, and improper storage. These toxins can cause a multitude of foodborne illnesses and can lead to cardiovascular aberration and adverse gastrointestinal symptoms. Analytical techniques are in use for the detection of toxins in fermented foods for monitoring and control purposes. These include culture, chromatographic, immunoassays, and nano sensor-based techniques. These detection techniques can be used during the production process and along the food chain. On an industrial scale, HPLC is widely used for sensitive quantification of toxins in fermented foods. Recently, biosensor and nano sensor-based techniques have gained popularity due to accuracy, time efficiency, and simultaneous detection of multiple toxins. Other strategic methods being investigated for the removal of toxins from fermented foods include the use of specific starter cultures for bio-preservation, aflatoxin-binding, and biogenic amine-degradation agents that may help to appropriately manage the food safety concerns associated with fermented foods.

Alternaria toxins are naturally occurring contaminants found in natural products. Given the prevalence of Alternaria toxins and the complexity of oil-rich matrices, achieving ultra-trace analysis has become a daunting task. A new sample pretreatment technique, i.e., cold-induced liquid-liquid microextraction combined with serially-coupled-columns for SIDA-UHPLC-MS/MS, was developed and reported for the first time. Theoretical and experimental investigations on the mechanism and key parameters revealed that the proposed method achieved simultaneous purification and enrichment in one-step sample extraction with a superior limit of quantitation (0.15-1.5 μg kg -1 ), without further sample manipulation, such as fat removal or solvent exchange procedures prior to LC-MS. The method was validated taking into consideration EU guidelines and showed acceptable linearity (r ≥ 0.9991), accuracy with recoveries between 75 and 114% and precision with RSD≤9.7% for all of the analytes studied. It was successfully applied to the analysis of twenty samples sourced from the Mediterranean region in order to gain first insights into Alternaria toxins contaminations in olive oils. This technical approach is well suited for large-scale studies in a high-throughput and cost-effective quality assurance laboratory environments, and it has the potential to detect ultra-trace levels of toxins in complex samples, which may lead to the development of new and sustainable sample preparation procedures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Lataste's viper (Vipera latastei) is a venomous European viper endemic to the Iberian Peninsula, recognised as medically important by the World Health Organization. To date, no comprehensive characterisation of this species' venom has been reported. Here, we analysed the venoms of juvenile and adult specimens of V. latastei from two environmentally different populations from northern Portugal. Using bottom-up venomics, we produced six venom proteomes (three per population) from vipers belonging to both age classes (i.e., two juveniles and four adults), and RP-HPLC profiles of 54 venoms collected from wild specimens. Venoms from juveniles and adults differed in their chromatographic profiles and relative abundances of their toxins, suggesting the occurrence of ontogenetic changes in venom composition. Specifically, snake venom metalloproteinase (SVMP) was the most abundant toxin family in juvenile venoms, while snake venom serine proteinases (SVSPs), phospholipases A 2 (PLA 2 s), and C-type lectin-like (CTLs) proteins were the main toxins comprising adult venoms. The RP-HPLC venom profiles were found to vary significantly between the two sampled localities, indicating geographic variability. Furthermore, the presence/absence of certain peaks in the venom chromatographic profiles appeared to be significantly correlated also to factors like body size and sex of the vipers. Our findings show that V. latastei venom is a variable phenotype. The intraspecific differences we detected in its composition likely mirror changes in the feeding ecology of this species, taking place during different life stages and under different environmental pressures. SIGNIFICANCE: Lataste's viper (Vipera latastei) is a medically important viper endemic to the Iberian Peninsula, inhabiting different habitats and undergoing a marked ontogenetic dietary shift. In the current study, we report the first proteomic analysis of V. latastei venom from two environmentally different localities in northern Portugal. Our bottom-up venomic analyses show that snake venom serine proteinases (SVSPs), phospholipases A2 (PLA2s), and C-type lectin-like (CTLs) proteins are the major components of adult V. latastei venom. The comparative analysis of young and adult venoms suggests the occurrence of ontogenetic shift in toxin abundances, with snake venom metalloproteinases (SVMPs) being the predominant toxins in juvenile venoms. Moreover, geographic venom variation between the two studied populations is also detected, with our statistical analyses suggesting that factors like body size and sex of the vipers are possibly at play in its determination. Our work represents the first assessment of the composition of V. latastei venom, and the first step towards a better understanding of the drivers behind its variability., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This focused review presents various approaches or formats in which liquid crystals (LCs) have been used as stimuli-responsive sensors. In these sensors, the LC molecules adopt some well-defined arrangement based on the sensor composition and the chemistry of the system. The sensor usually consists of a molecule or functionality in the system that engages in some form of specific interaction with the analyte of interest. The presence of analyte brings about the specific interaction, which then triggers an orientational transition of the LC molecules, which is optically discernible via a polarized optical image that shows up as dark or bright, depending on the orientation of the LC molecules in the system (usually a homeotropic or planar arrangement). The various applications of LCs as biosensors for glucose, protein and peptide detection, biomarkers, drug molecules and metabolites are extensively reviewed. The review also presents applications of LC-based sensors in the detection of heavy metals, anionic species, gases, volatile organic compounds (VOCs), toxic substances and in pH monitoring. Additionally discussed are the various ways in which LCs have been used in the field of material science. Specific attention has been given to the sensing mechanism of each sensor and it is important to note that in all cases, LC-based sensing involves some form of orientational transition of the LC molecules in the presence of a given analyte. Finally, the review concludes by giving future perspectives on LC-based sensors.

Consumers often believe that "natural food" is harmless, however naturally occurring toxins in food represent a health risk to humans. Honey as a natural, nutritious sweetener, is one of the most commonly consumed foods throughout the world. However, food safety concerns for honey arise when honeybees collect nectar from poisonous plants such as Rhododendron sp., Coriaria arborea, and Tripterygium wilfordii Hook F. Such honey contains natural plant toxins. Humans may develop intoxication symptoms after consuming toxic honey; in some cases, it can be fatal. As a result, toxic honey poses an often-ignored threat to public health. Typical plant toxins such as grayanotoxins, triptolides, tutin and pyrrolizidine alkaloids, have been identified in toxic honey. Although different toxic honeys elicit similar symptoms, such as vomiting, nausea, and dizziness, the mechanism of toxicity may be different. Thus, it is necessary to determine the exact toxicity mechanism of different toxins to further develop effective antidotes and cures. Another important challenge is preventing toxic honey from entering the food chain. Liquid chromatography-mass spectrometry has a wide range of applications in the detection of different toxins due to its accuracy and simplicity. More methods, however, are urgently needed to detect multiple plant-derived toxins in honey and its derivatives. Developing uniform international standards for toxin detection during quarantine using advanced techniques is critical for preventing human consumption of toxic honey., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The majority of the effects observed upon envenomation by scorpaenoid fish species can be reproduced by the cytolysins present in their venoms. Fish cytolysins are multifunctional proteins that elicit lethal, cytolytic, cardiovascular, inflammatory, nociceptive, and neuromuscular activities, representing a novel class of protein toxins. These large proteins (MW 150-320 kDa) are composed by two different subunits, termed α and β, with about 700 amino acid residues each, being usually active in oligomeric form. There is a high degree of similarity between the primary sequences of cytolysins from different fish species. This suggests these molecules share similar mechanisms of action, which, at least regarding the cytolytic activity, has been proved to involve pore formation. Although the remaining components of fish venoms have interesting biological activities, fish cytolysins stand out because of their multifunctional nature and their ability to reproduce the main events of envenomation on their own. Considerable knowledge about fish cytolysins has been accumulated over the years, although there remains much to be unveiled. In this review, we compiled and compared the current information on the biochemical aspects and pharmacological activities of fish cytolysins, going over their structures, activities, mechanisms of action, and perspectives for the future.

The presence of plant toxins and/or cyanotoxins in food supplements implies consumer health risks. Therefore, a targeted ultra-high performance liquid chromatographic-tandem mass spectrometric method to detect/quantify 25 toxins simultaneously in food supplement formulations was developed and validated. Full validation for tablets/powders and secondary validation for a liquid and soft gel capsule indicated that most compounds were efficiently extracted (≥ 75%), while others were only partly extracted (18 - 61%). Trueness was fulfilled (70 - 120%), with some exceptions (mostly at the lowest validation level). Intralaboratory repeatability, intra- and interlaboratory reproducibility values of ≤ 20%, ≤ 25%, and ≤ 25% were obtained for most, respectively. Matrix effects were found to be significant for most compounds. Good sensitivity (µg/kg level) was observed for galegin(e), lycopsamine, lycorine, rubiadin, skimmiamine, and vascin(e), in contrast to helveticoside, lucidin, lucidin-3-primveroside, plumbagin(e), and thujone, which were detected at the mg/kg level. The other compounds were characterized by a sensitivity between 10 to 1000 µg/kg. The validated methodology was applied for 52 food supplements (tablets, capsules, liquids/syrup, etc.) purchased from the Belgian market. In more than 25% of the samples, one or more toxins were detected (concentrations determined using standard addition). Lycopsamine, microcystin LR, solamargine, thujone, and vasicin(e) were the most frequently detected toxins. A clear link between the toxins detected and the plant species on the food supplement ingredient list could not always be established. This generic "dilute-and-shoot" procedure can be used for further research on toxins in food supplements and by extension other plant/algae-based food/feed commodities (herbs, edible flowers, etc.)., Competing Interests: The authors declare that they have no conflict of interest., (Thieme. All rights reserved.)

In the coastal countries of Southeast Asia, fish is a staple diet and certain fish species are food delicacies to local populations or commercially important to individual communities. Although there have been several suspected cases of ciguatera fish poisoning (CFP) in Southeast Asian countries, few have been confirmed by ciguatoxins identification, resulting in limited information for the correct diagnosis of this food-borne disease. In the present study, ciguatoxin-1B (CTX-1B) in red snapper ( Lutjanus bohar ) implicated in a CFP case in Sabah, Malaysia, in December 2017 was determined by single-quadrupole selected ion monitoring (SIM) liquid chromatography/mass spectrometry (LC/MS). Continuous consumption of the toxic fish likely resulted in CFP, even when the toxin concentration in the fish consumed was low. The identification of the fish species was performed using the molecular characterization of the mitochondrial cytochrome c oxidase subunit I gene marker, with a phylogenetic analysis of the genus Lutjanus . This is the first report identifying the causative toxin in fish-implicated CFP in Malaysia.

Acute kidney injury (AKI) is a significant risk factor for developing chronic kidney disease and progression to end-stage renal disease in elderly patients. AKI is also a relatively common complication after kidney transplantation (KTx) associated with graft failure. Since the lifespan of a transplanted kidney is limited, the risk of the loss/deterioration of graft function (DoGF) should be estimated to apply the preventive treatment. The collection of saliva and urine is more convenient than collecting blood and can be performed at home. The study aimed to verify whether non-invasive biomarkers, determined in saliva and urine, may be useful in the prediction of DoGF in kidney transplant recipients (KTRs) ( n = 92). Salivary and serum toxins ( p -cresol sulfate, pCS; indoxyl sulfate, IS) concentrations were determined using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Urinary proteins, hemoglobin, and glucose were measured using a semi-quantitative strip test. Salivary IS (odds ratio (OR) = 1.19), and proteinuria (OR = 3.69) were demonstrated as independent factors for the prediction of DoGF. Satisfactory discriminatory power (area under the receiver operating characteristic curve (AUC) = 0.71 ± 0.07) and calibration of the model were obtained. The model showed that categories of the increasing probability of the risk of DoGF are associated with the decreased risk of graft survival. The non-invasive diagnostic biomarkers are a useful screening tool to identify high-risk patients for DoGF.

Cereals and feed contaminated with ergot alkaloids (EAs) have been of concern for several decades. Nowadays, analysis of EAs is focused on ergometrine, ergotamine, ergosine, ergocristine, ergocryptine (a mixture of α- and β-isomers) and ergocornine and their related -inine epimers as listed in the European Commission Recommendation 2012/154/EU. Liquid chromatography with fluorescence detection has been used for quantification of EAs for decades whilst LC-MS has become the work-horse for quantification of EAs in the last decade. However, in LC-MS analysis matrix effects of different magnitudes exist for each EA epimer, especially ergometrine/ergometrinine, even after different clean-up procedures. This leads to an underestimation or overestimation of EAs levels. Moreover, isotopic labelled standards for EAs are still not available in the market. This review aims to provide background information on different analytical methods, discuss their advantages and disadvantages and possible advancement. Moreover, the method performance requirements to support forthcoming regulations are also discussed.

An analytical workflow including mass spectral library, generic sample preparation, chromatographic separation, and analysis by high-resolution mass spectrometry (HRMS) was developed to gain insight into the occurrence of plant toxins, mycotoxins and phytoestrogens in plant-based food. This workflow was applied to 156 compounds including 90 plant toxins (pyrrolizidine alkaloids, tropane alkaloids, glycoalkaloids, isoquinoline alkaloids and aristolochic acids), 54 mycotoxins (including ergot alkaloids and Alternaria toxins) and 12 phytoestrogens (including isoflavones, lignans and coumestan) in plant-based protein ingredients, cereal and pseudo-cereal products. A mass spectral library was built based on fragmentation spectra collected at 10 different collision energies in both positive and negative ionisation modes for each toxin. Emphasis was put on a generic QuEChERS-like sample preparation followed by ultra-high-pressure liquid chromatography using alkaline mobile phase allowing the separation of more than 50 toxic pyrrolizidine alkaloids. HRMS acquisition comprised a full-scan event for toxins detection followed by data-dependent MS2 for toxin identification against mass spectrum. Method performance was evaluated using fortified samples in terms of sensitivity, repeatability, reproducibility and recovery. All toxins were positively identified at levels ranging from 1 µg kg -1 to 100 µg kg -1 . Quantitative results obtained by a standard addition approach met SANTE/12682/2019 criteria for 132 out of 156 toxins. Such a workflow using generic, sensitive and selective multi-residue method allows a better insight into the occurrence of regulated and non-regulated toxins in plant-based foods and to conduct safety evaluation and risk assessments when needed.

The specificity and potency of venom components give them a unique advantage in developing various pharmaceutical drugs. Though venom is a cocktail of proteins, rarely are the synergy and association between various venom components studied. Understanding the relationship between various components of venom is critical in medical research. Using meta-analysis, we observed underlying patterns and associations in the appearance of the toxin families. For Crotalus , Dis has the most associations with the following toxins: PDE; BPP; CRL; CRiSP; LAAO; SVMP P-I and LAAO; SVMP P-III and LAAO. In Sistrurus venom, CTL and NGF have the most associations. These associations can predict the presence of proteins in novel venom and understand synergies between venom components for enhanced bioactivity. Using this approach, the need to revisit the classification of proteins as major components or minor components is highlighted. The revised classification of venom components is based on ubiquity, bioactivity, the number of associations, and synergies. The revised classification can be expected to trigger increased research on venom components, such as NGF, which have high biomedical significance. Using hierarchical clustering, we observed that the genera's venom compositions were similar, based on functional characteristics rather than phylogenetic relationships.

Safety and quality are key issues for the food industry. Consequently, there is growing demand to preserve the food chain and products against substances toxic, harmful to human health, such as contaminants, allergens, toxins, or pathogens. For this reason, it is mandatory to develop highly sensitive, reliable, rapid, and cost-effective sensing systems/devices, such as electrochemical sensors/biosensors. Generally, conventional techniques are limited by long analyses, expensive and complex procedures, and skilled personnel. Therefore, developing performant electrochemical biosensors can significantly support the screening of food chains and products. Here, we report some of the recent developments in this area and analyze the contributions produced by electrochemical biosensors in food screening and their challenges.

Phytotoxins are plant secondary metabolites. They have recently been considered as chemicals of emerging concern (CECs) and there is a growing interest in their environmental fate and potential threat to public health. Dedicated target and non-target screening (NTS) analysis of phytotoxins in environmental samples are sparse, meanwhile phytotoxins are rarely detected in NTS-based analysis due to lack of an efficient methodology. Development of new analytical measurement methods is therefore highly needed. In this study, we for the first time investigated key parameters of reversed phase liquid chromatography-high resolution mass spectrometry (RPLC-HRMS) for five major classes of phytotoxins (alkaloids, steroids, terpenoids, flavonoids and aromatic polyketides) in environmental matrices; the investigation included analytical conditions which have not yet been explored by others, e.g. ionization at alkaline pH above 9. As the outcome we established a new analytical method for target/suspect screening and NTS of phytotoxins in the environment, which significantly improved the detection sensitivity with up to 40 times compared to previous methods, and enabled the discovery of over 30 phytotoxins in a NTS-based environmental study. We also observed that the negative ionization of phenols could be facilitated by the number of hydroxyl groups on the ring rather than their position of substitution. This study is of interest for a better fundamental understanding of the behavior of phytotoxins in LC-MS. Dedicated target/suspect screening and NTS methods will facilitate a better risk characterization of phytotoxins in the environment and stimulate implementation of new public regulation on phytotoxins., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Aims: The aims of the study were to (i) improve the evaluation criteria of detoxifying Jatropha curcas L. cake (JCC), (ii) isolate and characterize a JCC tolerant strain, (iii) explore its JCC detoxifying potential., Methods and Results: The zebrafish was employed as a survival model to screen the strains capable of detoxifying JCC. A strain identified as Mucor circinelloides SCYA25, which is highly capable of degrading all toxic components, was isolated from soil. Different solid-state fermentation parameters were optimized by response surface methodology. The optimal values for inoculation amount, moisture content, temperature, and time were found to be 18% (1·8 × 10 6 spores g -1 cake), 66%, 26, and 36 days, respectively, to achieve maximum detoxification of the JCC (92%). Under optimal fermentation conditions, the protein content of JCC was increased, while the concentrations of ether extract, crude fiber, toxins, and anti-nutritional substances were all degraded considerably (P < 0·05). Scanning electron microscopy and Fourier transform infrared spectrometer analysis revealed that the fermentation process could disrupt the surface structure and improve the ratio of α-helix to β-folding in the JCC protein, which may improve the digestibility when the detoxified JCC is used as a feedstuff., Conclusions: Our results indicate that M. circinelloides SCYA25 is able to detoxify JCC and improve its nutritional profile, which is beneficial to the safe utilization of JCC as a protein feedstuff., Significance and Impact of the Study: The newly identified M. circinelloides SCYA25 detoxified JCC in a safe manner to provide a potential alternative to soybean meal for the feed industry. These results also provide a new perspective and method for the toxicity evaluation and utilization of JCC and similar toxic agricultural by-products., (© 2020 The Society for Applied Microbiology.)

Alternaria toxins are emerging mycotoxins whose regulation and standardization are in progress by the European Commission and the European Committee for Standardization. This paper describes a dilute and shoot approach to determine five Alternaria toxins in selected food samples using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The strategy involves sample extraction with acidified aqueous methanol, followed by a solvent change accomplished via sample evaporation and reconstitution. The quantification is based on isotope dilution, applying all corresponding isotopically labeled internal standards to compensate possible matrix effects of the analysis. The main advantages of the present method over other existing methods includes simple and effective sample preparation, as well as detection with high sensitivity. The five-fold sample dilution can decrease matrix effects, which were evaluated with both external and internal standard methods. The results demonstrated a limit of quantification lower than 1.0 µg/kg for all five analytes for the first time. The newly presented method showed acceptable accuracy (52.7-111%) when analyzing naturally contaminated and spiked standard samples at the described levels. The method was validated for tomato-based and flour samples (wheat, rye, and maize). The absolute recovery ranged from 66.7% to 91.6% (RSD < 10%). The developed method could be an alternative approach for those laboratories that exclude sample cleanup and pre-concentration of state-of-the-art instruments with enhanced sensitivity.

Conventional absorbents for hemoperfusions suffer from low efficiency and slow absorption with numerous side effects. In this research, we developed cellulose acetate (CA) functionalized graphene oxide (GO) beads (∼1.5-2 mm) that can be used for direct hemoperfusion, aiming at the treatment of kidney dysfunction. The CA-functionalized GO bead facilitates adsorption of toxins with high biocompatibility and high-efficiency of hemoperfusion while maintaining high retention for red blood cell, white blood cells, and platelets. Our in vitro results show that the toxin concentration for creatinine reduced from 0.21 to 0.12 μM ( p < 0.005), uric acid from 0.31 to 0.15 mM ( p < 0.005), and bilirubin from 0.36 to 0.09 mM ( p < 0.005), restoring to normal levels within 2 h. Our in vivo study on rats (Sprague-Dawley, n = 30) showed that the concentration for creatinine reduced from 83.23 to 54.87 μmol L -1 ( p < 0.0001) and uric acid from 93.4 to 54.14 μmol L -1 ( p < 0.0001), restoring to normal levels within 30 min. Results from molecular dynamics (MD) simulations using free-energy calculations reveal that the presence of CA on GO increases the surface area for adsorption and enhances penetration of toxins in the binding cavities because of the increased electrostatic and van der Waals force (vdW) interactions. These results provide critical insight to fabricate graphene-based beads for hemoperfusion and to have the potential for the treatment of blood-related disease.

Hydrogels are three-dimensional porous polymeric networks prepared by physical or chemical cross-linking of hydrophilic molecules, which can be made into smart materials through judicious chemical modifications to recognize external stimuli; more specifically, this can be accomplished by the integration with stimuli-responsive polymers or sensing molecules that has drawn considerable attention in their possible roles as sensors and diagnostic tools. They can be tailored in different structures and integrated into systems, depending on their chemical and physical structure, sensitivity to the external stimuli and biocompatibility. A panoramic overview of the sensing advances in the field of hydrogels over the past several decades focusing on a variety protocols of hydrogel preparations is provided, with a major focus on natural polymers. The modifications of hydrogel composites by incorporating inorganic nanoparticles and organic polymeric compounds for sensor applications and their mechanisms are also discussed.

The toxin abrin found in the seeds of Abrus precatorius has attracted much attention regarding criminal and terroristic misuse over the past decade. Progress in analytical methods for a rapid and unambiguous identification of low abrin concentrations in complex matrices is essential. Here, we report on the development and evaluation of a MALDI-TOF mass spectrometry approach for the fast, sensitive and robust abrin isolectin identification, differentiation and quantification in complex food matrices. The method combines immunoaffinity-enrichment with specific abrin antibodies, accelerated trypsin digestion and the subsequent MALDI-TOF analysis of abrin peptides using labeled peptides for quantification purposes. Following the optimization of the workflow, common and isoform-specific peptides were detected resulting in a ~38% sequence coverage of abrin when testing ng-amounts of the toxin. The lower limit of detection was established at 40 ng/mL in milk and apple juice. Isotope-labeled versions of abundant peptides with high ionization efficiency were added. The quantitative evaluation demonstrated an assay variability at or below 22% with a linear range up to 800 ng/mL. MALDI-TOF mass spectrometry allows for a simple and fast (<5 min) analysis of abrin peptides, without a time-consuming peptide chromatographic separation, thus constituting a relevant alternative to liquid chromatography-tandem mass spectrometry.

Venomics, the study of biological venoms, could potentially provide a new source of therapeutic compounds, yet information on the venoms from marine organisms, including cnidarians (sea anemones, corals, and jellyfish), is limited. This study identified the putative toxins of two species of jellyfish-edible jellyfish Rhopilema esculentum Kishinouye, 1891, also known as flame jellyfish, and Amuska jellyfish Sanderia malayensis Goette, 1886. Utilizing nano-flow liquid chromatography tandem mass spectrometry (nLC-MS/MS), 3000 proteins were identified from the nematocysts in each of the above two jellyfish species. Forty and fifty-one putative toxins were identified in R. esculentum and S. malayensis , respectively, which were further classified into eight toxin families according to their predicted functions. Amongst the identified putative toxins, hemostasis-impairing toxins and proteases were found to be the most dominant members (>60%). The present study demonstrates the first proteomes of nematocysts from two jellyfish species with economic and environmental importance, and expands the foundation and understanding of cnidarian toxins.

The evolution of instrumentation in terms of separation and detection allowed a real improvement of the sensitivity and analysis time. However, the analysis of ultra-traces of toxins in complex samples requires often a step of purification and even preconcentration before their chromatographic analysis. Therefore, immunoaffinity sorbents based on specific antibodies thus providing a molecular recognition mechanism appear as powerful tools for the selective extraction of a target molecule and its structural analogs to obtain more reliable and sensitive quantitative analysis in environmental, food or biological matrices. This review focuses on immunosorbents that have proven their efficiency in selectively extracting various types of toxins of various sizes (from small mycotoxins to large proteins) and physicochemical properties. Immunosorbents are now commercially available, and their use has been validated for numerous applications. The wide variety of samples to be analyzed, as well as extraction conditions and their impact on extraction yields, is discussed. In addition, their potential for purification and thus suppression of matrix effects, responsible for quantification problems especially in mass spectrometry, is presented. Due to their similar properties, molecularly imprinted polymers and aptamer-based sorbents that appear to be an interesting alternative to antibodies are also briefly addressed by comparing their potential with that of immunosorbents.

Mycotoxins are common in grains in sub-Saharan Africa and negatively impact human and animal health and production. This study assessed occurrences of mycotoxins, some plant, and bacterial metabolites in 16 dairy and 27 poultry feeds, and 24 feed ingredients from Machakos town, Kenya, in February and August 2019. We analyzed the samples using a validated multi-toxin liquid chromatography-tandem mass spectrometry method. A total of 153 mycotoxins, plant, and bacterial toxins, were detected in the samples. All the samples were co-contaminated with 21 to 116 different mycotoxins and/or metabolites. The commonly occurring and EU regulated mycotoxins reported were; aflatoxins (AFs) (70%; range 0.2-318.5 μg/kg), deoxynivalenol (82%; range 22.2-1037 μg/kg), ergot alkaloids (70%; range 0.4-285.7 μg/kg), fumonisins (90%; range 32.4-14,346 μg/kg), HT-2 toxin (3%; range 11.9-13.8 μg/kg), ochratoxin A (24%; range 1.1-24.3 μg/kg), T-2 toxin (4%; range 2.7-5.2 μg/kg) and zearalenone (94%; range 0.3-910.4 μg/kg). Other unregulated emerging mycotoxins and metabolites including Alternaria toxins, Aspergillus toxins, bacterial metabolites, cytochalasins, depsipeptides, Fusarium metabolites, metabolites from other fungi, Penicillium toxins, phytoestrogens, plant metabolites, and unspecific metabolites were also detected at varying levels. Except for total AFs, where the average contamination level was above the EU regulatory limit, all the other mycotoxins detected had average contamination levels below the limits. Ninety-six percent of all the samples were contaminated with more than one of the EU regulated mycotoxins. These co-occurrences may cause synergistic and additive health effects thereby hindering the growth of the Kenyan livestock sector.

This study presents the application of a suspect screening approach to screen a wide range of natural toxins, including mycotoxins, bacterial toxins, and plant toxins, in surface waters. The method is based on a generic solid-phase extraction procedure, using three sorbent phases in two cartridges that are connected in series, hence covering a wide range of polarities, followed by liquid chromatography coupled to high-resolution mass spectrometry. The acquisition was performed in the full-scan and data-dependent modes while working under positive and negative ionisation conditions. This method was applied in order to assess the natural toxins in the Ter River water reservoirs, which are used to produce drinking water for Barcelona city (Spain). The study was carried out during a period of seven months, covering the expected prior, during, and post-peak blooming periods of the natural toxins. Fifty-three (53) compounds were tentatively identified, and nine of these were confirmed and quantified. Phytotoxins were identified as the most frequent group of natural toxins in the water, particularly the alkaloids group. Finally, the toxins identified to levels 2 and 1 were prioritised according to their bioaccumulation factor, biodegradability, frequency of detection, and toxicity. This screening and prioritisation approach resulted in different natural toxins that should be further assessed for their ecotoxicological effects and considered in future studies.

Biological toxins are a heterogeneous group of high molecular as well as low molecular weight toxins produced by living organisms. Due to their physical and logistical properties, biological toxins are very attractive to terrorists for use in acts of bioterrorism. Therefore, among the group of biological toxins, several are categorized as security relevant, e.g., botulinum neurotoxins, staphylococcal enterotoxins, abrin, ricin or saxitoxin. Additionally, several security sensitive toxins also play a major role in natural food poisoning outbreaks. For a prompt response to a potential bioterrorist attack using biological toxins, first responders need reliable, easy-to-use and highly sensitive methodologies for on-site detection of the causative agent. Therefore, the aim of this review is to present on-site immunoassay platforms for multiplex detection of biological toxins. Furthermore, we introduce several commercially available detection technologies specialized for mobile or on-site identification of security sensitive toxins.

Accumulation of protein-bound uremic toxins (PBUTs) has a high incidence in the blood of hemodialysis-treated patients with chronic kidney disease. Development of adsorbent for the high-efficient, selective, and removal of various PBUTs is still challenging because of their strong interactions to the corresponding binding proteins and the complex blood background. Herein, we reported poly-cyclodextrins adsorbents with multi adsorption-sites for PBUTs removal from plasma. Compared to poly-α-cyclodextrin and poly-γ-cyclodextrin, poly-β-cyclodextrin (PβCD) showed the best adsorption capability and the maximum p-cresol sulfate (PCS, a model PBUT) binding capacity of PβCD (263 mg g -1 ). Combining with hemodialysis, PβCD adsorbents added into the dialysate can remove 96 % PCS in the plasma via adsorbent once-through mode. Additionally, various PBUTs co-exsiting in the plasma could be effectively removed, exhibiting high-concentrition PBUT adsorption property of PβCD. We expect that the PCD adsorbents combining with hemodialysis therapy may become a promising potential for clinical PBUT removal., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The first quantitative multiclass approach enabling the accurate quantification of >1200 biotoxins, pesticides and veterinary drugs in complex feed using liquid chromatography tandem mass spectrometry (LC-MS/MS) has been developed. Optimization of HPLC/UHPLC (chromatographic column, flow rate and injection volume) and MS/MS conditions (dwell time and cycle time) were carried out in order to allow the combination of five major substance classes and the high number of target analytes with different physico-chemical properties. Cycle times and retention windows were carefully optimized and ensured appropriate dwell times reducing the overall measurement error. Validation was carried out in two compound feed matrices according to the EU SANTE validation guideline. Apparent recoveries matching the acceptable range of 60-140% accounted 60% and 79% for all analytes in cattle and chicken feed, respectively. High extraction efficiencies were obtained for all analyte/matrix combinations and revealed matrix effects as the main source for deviation of the targeted performance criteria. Concerning the methods repeatability 99% of all analytes in chicken and 96% in cattle feed complied with the acceptable RSD ≤ 20% criterion. Limits of quantification were between 1-10 µg/kg for the vast majority of compounds. Finally, the methods applicability was tested in >130 real compound feed samples and provides first insights into co-exposure of agro-contaminants in animal feed., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interest., (Copyright © 2020. Published by Elsevier B.V.)

Nanobioconjugates are hybrid materials that result from the coalescence of biomolecules and nanomaterials. They have emerged as a strategy to amplify the signal response in the biosensor field with the potential to enhance the sensitivity and detection limits of analytical assays. This critical review collects a myriad of strategies for the development of nanobioconjugates based on the conjugation of proteins, antibodies, carbohydrates, and DNA/RNA with noble metals, quantum dots, carbon- and magnetic-based nanomaterials, polymers, and complexes. It first discusses nanobioconjugates assembly and characterization to focus on the strategies to amplify a biorecognition event in biosensing, including molecular-, enzymatic-, and electroactive complex-based approaches. It provides some examples, current challenges, and future perspectives of nanobioconjugates for the amplification of signals in electrochemical biosensing.

Tall fescue (Lolium arundinaceum) is a widely used forage grass which shares a symbiosis with the endophytic fungus Epichloë coenophiala. The endophyte produces an alkaloid toxin that provides herbivory, heat and drought resistance to the grass, but can cause fescue toxicosis in grazing livestock. Fescue toxicosis can lead to reduced weight gain and milk yields resulting in significant losses to the livestock industry. The objective of this study was to identify bacterial and fungal communities associated with fescue toxicosis tolerance. In this trial, 149 Angus cows across two farms were continuously exposed to toxic, endophyte-infected, fescue for a total of 13 weeks. Of those 149 cows, 40 were classified into either high (HT) or low (LT) tolerance groups according to their growth performance (weight gain). 20 HT and 20 LT cattle balanced by farm were selected for amplicon sequencing to compare the fecal microbiota of the two tolerance groups. This study reveals significantly (q<0.05) different bacterial and fungal microbiota between HT and LT cattle, and indicates that fungal phylotypes may be important for an animal's response to fescue toxicosis: We found that fungal phylotypes affiliating to the Neocallimastigaceae, which are known to be important fiber-degrading fungi, were consistently more abundant in the HT cattle. Whereas fungal phylotypes related to the genus Thelebolus were more abundant in the LT cattle. This study also found more pronounced shifts in the microbiota in animals receiving higher amounts of the toxin. We identified fungal phylotypes which were consistently more abundant either in HT or LT cattle and may thus be associated with the respective animal's response to fescue toxicosis. Our results thus suggest that some fungal phylotypes might be involved in mitigating fescue toxicosis., Competing Interests: The authors have declared that no competing interests exist.

Food, by nature, is a biological substrate and is therefore capable of supporting the growth of microbials that are potential producers of toxic compounds [...].

Toxins are one of the major threatening factors to human and animal health, as well as economic growth. There is therefore an urgent demand from various communities to develop novel analytical methods for the sensitive detection of toxins in complex matrixes. Among the as-developed toxin detection strategies, nanocomposite-based aptamer sensors (termed as aptasensors) show tremendous potential for combating toxin pollution; in particular electrochemical (EC) aptasensors have received significant attention because of their unique advantages, including simplicity, rapidness, high sensitivity, low cost and suitability for field-testing. This paper reviewed the recently published approaches for the development of nanocomposite-/nanomaterial-based EC aptasensors for the detection of toxins with high assaying performance, and their potential applications in environmental monitoring, clinical diagnostics, and food safety control by summarizing the detection of different types of toxins, including fungal mycotoxins, algal toxins and bacterial enterotoxins. The effects of nanocomposite properties on the detection performance of EC aptasensors have been fully addressed for supplying readers with a comprehensive understanding of their improvement. The current technical challenges and future prospects of this subject have also been discussed.

The purpose of this study was to set up a sensitive method for the simultaneous determination of phycotoxins and cyanotoxins-Emerging pollutants with different structures and harmful properties (hepatotoxicity, neurotoxicity and cytotoxicity)-In environmental waters. Due to the low concentrations detected in these samples, a pre-concentration step is required and here it was performed in a single step with a commercial cartridge (Strata™-X), achieving enrichment factors up to 200 and satisfactory recovery (R = 70-118%) in different aqueous matrices. After solid-phase extraction (SPE), toxins were separated and quantified by High Performance Liquid Chromatography- Heated ElectroSpray Ionisation Tandem Mass Spectrometry (HPLC-HESI-MS/MS) in Multiple Reaction Monitoring (MRM) mode. An analytical evaluation of the proposed method was done based on the analytical figures of merit, such as precision and trueness, linearity, selectivity, and sensitivity, and it turned out to be a robust tool for the quantification of ng L -1 levels, phycotoxins and cyanotoxins in both freshwater and saltwater samples.

Convection-based renal replacement therapies (RRTs) have the potential to improve patient outcomes when compared to diffusion-based RRT such as hemodialysis (HD), but have limited clearance rates. We propose and characterize multipoint dilution hemofiltration (MPD-HF), a purely convective blood purification technology which removes the fundamental filtration limit associated with convective RRT resulting in clearance rates on par with HD. In MPD-HF, filtration of liquid and solutes occurs along the length of the hollow fibers that convey the blood, and substitution fluid is pushed into the fibers at multiple points along their length. Since multiple filtration and dilution steps are contained within one pass of the blood through the hollow fiber, the fraction of fluid that can be filtered may be increased to allow a high clearance rate that removes a wide range of toxins. In vitro tests yielded an average steady-state filtrate fraction of 68%, exceeding commercial HDF cartridge filtrate fractions by a factor of approximately 3. The molecular weights of molecules cleared spans up to the cutoff of 66 kDa for albumin., (© 2020 International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.)

Protein biomarkers often exist as degradation fragments in biological samples, and affinity agents derived using a purified protein may not recognize them, limiting their value for clinical diagnosis. Herein, we present a method to overcome this issue, by selecting aptamers against a degraded form of the toxin B protein, which is a marker for diagnosing toxigenic Clostridium difficile infections. This approach has led to isolation of a DNA aptamer that recognizes degraded toxin B, fresh toxin B, and toxin B spiked into human stool samples. DNA aptamers selected using intact recombinant toxin B failed to recognize degraded toxin B, which is the form present in stored stool samples. Using this new aptamer, we produced a simple paper-based analytical device for colorimetric detection of toxin B in stool samples, or in the NAP1 strain of Clostridium difficile. The combined aptamer-selection and paper-sensing strategy can expand the practical utility of DNA aptamers in clinical diagnosis., (© 2020 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Viscum album lectin 1 (Viscumin) is one of the most important plant-based protein of potential adjuvant in cancer treatment. Therefore, the use of nano-biosensor technology as a novel emerges of biosensors is crucial to detect this modal agent in pharmacological study. Molecular imprinted polymer using 9-mer peptides sequence (epitope) was applied as a template. Using ultraviolet light, hydrogen bonding attained between the functional monomer and epitope, leading to the formation of a molecularly imprinted polymer. In the following, the epitope was derived from the surface of the polymer by sodium dodecyl sulfate (SDS) 2.5% and acetic acid 0.6% w/w. Finally, the designed nano-biosensor was exposed to different concentrations of the epitope. The selectivity of the nano-biosensor was tested in complex matrices such as blood plasma and urine. The scatchard analysis was covered for a consequence of the dissociation constants and the numbers of binding sites. Based on the results, the designed nano-biosensor has a limit of detection of 0.117 ng/μl and limit of quantification of 0.517 ng/μl in PBS buffer, respectively. These amounts stood 0.5 ng/μl and 0.8 ng/μl for urine environment and 1.25 ng/μl and 5 ng/μl for human blood fresh frozen plasma in the presence of ricin as the most homologue of viscumin (ML1) in fixed concentration (12:1), respectively. The time of detection and optimum pH was 8.0 min and 7.4, respectively. Designed and synthesized nano-biosensor is adequately qualified to be used in diverse complex areas, due to good efficiency., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)