Toxin In Food Samples Research Articles

Recent Advances in Monitoring Microbial Toxins in Food Samples by HPLC-Based Techniques: A Review

This study examines the significant impact of bacterial, algal, and fungal toxins on foodborne illnesses, and stresses the importance of advanced detection techniques, such as high-performance liquid chromatography (HPLC)-based methodologies. It emphasizes the urgent need for further advancements in these techniques to ensure food safety, as they offer significant benefits, including low detection limits and the ability to be combined with other techniques to detect a wide range of toxins. In this regard, HPLC has emerged as a versatile and sensitive analytical technique for this purpose. Various HPLC methods, often enhanced with detectors such as ultraviolet (UV), fluorescence (FD), and mass spectrometry (MS), have been developed to identify and quantify microbial toxins in a wide variety of food samples. Recent advancements include HPLC-FD methods that utilize the natural fluorescence of certain aflatoxins, improving detection sensitivity. HPLC-MS/MS and UHPLC-MS/MS techniques offer high selectivity and sensitivity, making them suitable for detecting a wide range of toxins in trace quantities. The adaptability of HPLC, combined with innovative detection technologies and sample preparation methods, holds significant potential for enhancing food safety monitoring and reducing the global burden of foodborne diseases.

Specific and Simultaneous Detection of E. coli O157:H7 and Shiga-like Toxins Using a Label-Free Photonic Immunosensor

The current study outlines the advancement of an innovative technique for the simultaneous detection of E. coli O157:H7 and its Shiga-like toxins in food samples by utilizing a photonic label-free biosensor coupled with a microfluidic system. This detection method relies on ring resonator transduction that is functionalized with specific bioreceptors against O157:H7 on silicon nitride surfaces capable of binding specifically to the antigen bacterium and its verotoxins. This experiment included the characterization of selected monoclonal and polyclonal antibodies employed as detection probes through ELISA immunoassays exposed to target bacterial antigens. A thorough validation of photonic immunosensor detection was conducted on inoculated minced beef samples using reference standards for E. coli O157:H7 and its verotoxins (VTx1 and VTx2) and compared to gold-standard quantification. The lowest limit-of-detection values of 10 CFU/mL and 1 ppm were achieved for the detection of bacteria and its verotoxins. In this study, the lowest limit of quantification (LoQ) achieved for bacterial quantification was 100 CFU/mL, and, for verotoxins, it was 2 ppm. This study confirmed the effectiveness of a new quality control and food hygiene method, demonstrating the rapid and sensitive detection of E. coli O157:H7 and its verotoxins. This innovative approach has the potential to be applied in food production environments.

Recent Advances in the Detection of Food Toxins Using Mass Spectrometry.

Edibles are the only source of nutrients and energy for humans. However, ingredients of edibles have undergone many physicochemical changes during preparation and storage. Aging, hydrolysis, oxidation, and rancidity are some of the major changes that not only change the native flavor, texture, and taste of food but also destroy the nutritive value and jeopardize public health. The major reasons for the production of harmful metabolites, chemicals, and toxins are poor processing, inappropriate storage, and microbial spoilage, which are lethal to consumers. In addition, the emergence of new pollutants has intensified the need for advanced and rapid food analysis techniques to detect such toxins. The issue with the detection of toxins in food samples is the nonvolatile nature and absence of detectable chromophores; hence, normal conventional techniques need additional derivatization. Mass spectrometry (MS) offers high sensitivity, selectivity, and capability to handle complex mixtures, making it an ideal analytical technique for the identification and quantification of food toxins. Recent technological advancements, such as high-resolution MS and tandem mass spectrometry (MS/MS), have significantly improved sensitivity, enabling the detection of food toxins at ultralow levels. Moreover, the emergence of ambient ionization techniques has facilitated rapid in situ analysis of samples with lower time and resources. Despite numerous advantages, the widespread adoption of MS in routine food safety monitoring faces certain challenges such as instrument cost, complexity, data analysis, and standardization of methods. Nevertheless, the continuous advancements in MS-technology and its integration with complementary techniques hold promising prospects for revolutionizing food safety monitoring. This review discusses the application of MS in detecting various food toxins including mycotoxins, marine biotoxins, and plant-derived toxins. It also explores the implementation of untargeted approaches, such as metabolomics and proteomics, for the discovery of novel and emerging food toxins, enhancing our understanding of potential hazards in the food supply chain.

Nanoscale Materials Applying for the Detection of Mycotoxins in Foods.

Trace amounts of mycotoxins in food matrices have caused a very serious problem of food safety and have attracted widespread attention. Developing accurate, sensitive, rapid mycotoxin detection and control strategies adapted to the complex matrices of food is crucial for in safeguarding public health. With the continuous development of nanotechnology and materials science, various nanoscale materials have been developed for the purification of complex food matrices or for providing response signals to achieve the accurate and rapid detection of various mycotoxins in food products. This article reviews and summarizes recent research (from 2018 to 2023) on new strategies and methods for the accurate or rapid detection of mold toxins in food samples using nanoscale materials. It places particular emphasis on outlining the characteristics of various nanoscale or nanostructural materials and their roles in the process of detecting mycotoxins. The aim of this paper is to promote the in-depth research and application of various nanoscale or structured materials and to provide guidance and reference for the development of strategies for the detection and control of mycotoxin contamination in complex matrices of food.

Egy LC-MS/MS alapú élelmiszervizsgálati módszer nemzetközi szabványosítása: egységesen elfogadott vizsgálati eljárás kidolgozása Alternaria toxinokra

There are more than seventy varieties of Alternaria toxins, but researchers have so far identified only a few of them structurally. The objective of this paper is to present a nearly ten-year process, during which an international standard for the simultaneous analysis of five Alternaria toxins in food samples was developed. This long process includes the development of the need for the standard and, in addition to the preparation and evaluation of the standardization tender, the development of the method, its validation and documentation. The paper focuses mainly on the development and validation of the analytical method, which is the longest and most labor-intensive part of the process, but in order to understand the overall picture, it is also necessary to emphasize the first and final steps. The development of a standard is a task of great responsibility for both the preparers of the standard and those involved in the validation and documentation of the standard, as the use of standardized methods is expected by the customers of the laboratories. On the other hand, laboratories that choose unique, self-developed methods can ascertain the accuracy and precision of their procedure by comparing them with the standard method. In this process that went on for nearly ten years, the original analytical method underwent several changes; the goal of these improvements was to make the procedure as simple and reproducible as possible. This is how the use isotope dilution mass spectrometry was reached through derivatization. It is important to emphasize that one of the goals of standardization is to have an appropriate analytical method available to authority laboratories for the testing of legally prescribed food contaminants, which procedure is available to any laboratory, however, it is questionable, whether the cost of the test covers its application. Consequently, it is not necessarily the most cost-effective analysis which is recommended by the standard, which may be the cause of conflict between the professional and economic managers of a laboratory in the case of private laboratories. The final form of the liquid chromatography/isotope dilution mass spectrometry (LC-IDMS) standard method developed for Alternaria toxins is likely to be approved and published by the European Committee for Standardization (CEN) in the end of 2021 (the standard has been issued since the article was submitted: CEN EN 17521:2021 Foodstuffs - Determination of Alternaria toxins in tomato, wheat and sunflower seeds by SPE clean-up and HPLC-MS/MS. The Editor). The standard will contain the determination of tenuazonic acid (TEA), altenuane (ALT), alternariol (AOH), tentoxin (TEN) and alternariol monomethyl ether (AME).

A novel colorimetric biosensor for sensitive detection of aflatoxin mediated by bacterial enzymatic reaction in saffron samples

Aflatoxin is regarded as the potent carcinogenic agent which is secreted from fungi and present in some food products. So far, many detection methods have been developed to determine the trace amounts of aflatoxin in foods. In the present study a colorimetric competitive assay for detection of aflatoxin B1 (AFB1) has been developed based on interaction of gelatin functionalized gold nanoparticles (AuNPs@gelatin) in specific enzymatic reaction. Bacterial supernatant containing gelatinase enzyme were used as the substrate that could digest the coated gelatin on the surface of AuNPs and following in the presence of NaCl medium ingredient resulted to color change of AuNPs colloidal solution from red to purple. It was observed that with addition of aflatoxin to the bacterial supernatant, aflatoxin could interfere in aggregation of AuNPs and inhibited the process which subsequently prevent the expected color change induced by AuNPs aggregation. The supernatant containing AuNPs were investigated to analyze their induced surface plasmon resonance spectra through UV–visible spectroscopy. The absorption values were directly proportional with the applied AFB1 concentration. The experiment conditions including incubation time, AuNPs concentration and pH were investigated. The obtained results showed that through this approach we could detect the AFB1 in a linear range from 10 to 140 pg ml−1, with detection limit of 4 pg ml−1. Real sample assay in saffron samples showed recoveries percentage of 92.4%–95.3%. The applied approach proposed simple, cost effective and specific method for detection of AFB1 toxin in food samples.

Detection of toxins involved in foodborne diseases caused by Gram-positive bacteria.

Bacterial toxins are food safety hazards causing about 10% of all reported foodborne outbreaks in Europe. Pertinent to Gram-positive pathogens, the most relevant toxins are emetic toxin and diarrheal enterotoxins of Bacillus cereus, neurotoxins of Clostridium botulinum, enterotoxin of Clostridium perfringens, and a family of enterotoxins produced by Staphylococcus aureus and some other staphylococci. These toxins are the most important virulence factors of respective foodborne pathogens and a primary cause of the related foodborne diseases. They are proteins or peptides that differ from each other in their size, structure, toxicity, toxicological end points, solubility, and stability, types of food matrix to which they are mostly related to. These differences influence the characteristics of required detection methods. Therefore, detection of these toxins in food samples, or detection of toxin production capacity in the bacterial isolate, remains one of the cornerstones of microbial food analysis and an essential tool in understanding the relevant properties of these toxins. Advanced research has led into new insights of the incidence of toxins, mechanisms of their production, their physicochemical properties, and their toxicological mode of action and dose-response profile. This review focuses on biological, immunological, mass spectrometry, and molecular assays as the most commonly used detection and quantification methods for toxins of B.cereus, C.botulinum, C.perfringens, and S. aureus. Gathered and analyzed information provides a comprehensive blueprint of the existing knowledge on the principles of these assays, their application in food safety, limits of detection and quantification, matrices in which they are applicable, and type of information they provide to the user.

New Advanced Materials and Sorbent-Based Microextraction Techniques as Strategies in Sample Preparation to Improve the Determination of Natural Toxins in Food Samples.

Natural toxins are chemical substances that are not toxic to the organisms that produce them, but which can be a potential risk to human health when ingested through food. Thus, it is of high interest to develop advanced analytical methodologies to control the occurrence of these compounds in food products. However, the analysis of food samples is a challenging task because of the high complexity of these matrices, which hinders the extraction and detection of the analytes. Therefore, sample preparation is a crucial step in food analysis to achieve adequate isolation and/or preconcentration of analytes and provide suitable clean-up of matrix interferences prior to instrumental analysis. Current trends in sample preparation involve moving towards “greener” approaches by scaling down analytical operations, miniaturizing the instruments and integrating new advanced materials as sorbents. The combination of these new materials with sorbent-based microextraction technologies enables the development of high-throughput sample preparation methods, which improve conventional extraction and clean-up procedures. This review gives an overview of the most relevant analytical strategies employed for sorbent-based microextraction of natural toxins of exogenous origin from food, as well as the improvements achieved in food sample preparation by the integration of new advanced materials as sorbents in these microextraction techniques, giving some relevant examples from the last ten years. Challenges and expected future trends are also discussed.

Magnetic quantum dot based lateral flow assay biosensor for multiplex and sensitive detection of protein toxins in food samples

Protein toxins, such as botulinum neurotoxin type A (BoNT/A) and staphylococcal enterotoxin B (SEB), easily pollute food and water and are ultra-toxic to humans and animals, thus requiring a sensitive on-site detection method. In this study, we reported a novel lateral flow assay (LFA) strip on the basis of magnetic quantum dot nanoparticles (MagQD NPs) for sensitive and multiplex protein toxin detection in food samples. A new type of MagQD NP was prepared by fixing the dense carboxylated QDs on the surface of polyethyleneimine-modified Fe3O4 magnetic NPs (MNPs) and applied in LFA with the following functions: capture and enrich target toxins from sample solutions and serve as advanced fluorescent labels for the quantitative determination of targets on the strip. Through this strategy, the assay realized quantified BoNT/A and SEB detection in 30 min with the limits of detection of 2.52 and 2.86 pg/mL, respectively. The selectivity and the ability of quantitative analysis of the method were validated in real food samples, including milk and juice. This MagQD-LFA biosensor showed considerable potential as a point-of-care testing tool for the sensitive detection of trace toxins.

Sensitive Monitoring of Enterobacterial Contamination of Food Using Self-Propelled Janus Microsensors.

Food poisoning caused by bacteria is a major cause of disease and death worldwide. Herein we describe the use of Janus micromotors as mobile sensors for the detection of toxins released by enterobacteria as indicators of food contamination. The micromotors are prepared by a Pickering emulsion approach and rely on the simultaneous encapsulation of platinum nanoparticles for enhanced bubble-propulsion and receptor-functionalized quantum dots (QDs) for selective binding with the 3-deoxy-d-manno-oct-2-ulosonic acid target in the endotoxin molecule. Lipopolysaccharides (LPS) from Salmonella enterica were used as target endotoxins, which upon interaction with the QDs induce a rapid quenching of the native fluorescence of the micromotors in a concentration-dependent manner. The micromotor assay can readily detect concentrations as low as 0.07 ng mL-1 of endotoxin, which is far below the level considered toxic to humans (275 μg mL-1). Micromotors have been successfully applied for the detection of Salmonella toxin in food samples in 15 min compared with several hours required by the existing Gold Standard method. Such ultrafast and reliable approach holds considerable promise for food contamination screening while awaiting the results of bacterial cultures in a myriad of food safety and security defense applications.

Use of T-2 toxin-immobilized amine-activated beads as an efficient affinity purification matrix for the isolation of specific IgY

An affinity purification method that isolates T-2 toxin-specific IgY utilizing a T-2-toxin-immobilized column was developed. The T-2 toxin was covalently coupled via a carbonyldiimidazole-activated hydroxyl functional group to amine-activated sepharose beads. The affinity-purified IgY was characterized by gel electrophoresis, fast protein liquid chromatography, enzyme-linked immunosorbant assay, surface plasmon resonance and mass spectrometry. A competitive inhibition ELISA (CI-ELISA) was performed using affinity-purified IgY with a T-2 toxin detection sensitivity of 30 ng/mL, which falls within the maximum permissible limit of 100 ng/mL. The cross reactivity of IgY towards deoxynivalenol, zearalenone, fumonisin B1 and HT-2 was significantly reduced after affinity purification. A surface plasmon resonance (SPR)-based inhibition assay was also applied for quantitative determination of T-2 toxin in spiked wheat samples. The results obtained indicate the feasibility of utilizing this IgY-based assay for the detection of T-2 toxin in food samples.

Development of colloidal gold-based immunochromatographic assay for the rapid detection of ricin toxin in food samples

A rapid, sensitive immunochromatographic (IC) assay was developed to detect ricin toxin in assay buffer and food samples. The assay was based on the sandwich format using a monoclonal antibody (mAb) and a polyclonal antibody (pAb) of two distinct specificities. The ricin-containing sample was added to the membrane and allowed to react with pAb-coated particles. The mixture was then passed along the porous membrane by capillary action past the mAb in the detection zone, which would bind the particles that had ricin bound to their surfaces, giving a red colour within the detection zone with an intensity proportional to the ricin concentration. In the absence of ricin, no immunogold was bound to the solid-phase antibody. With this method, 30 ng/ml of ricin toxin was detected in less than 10 min, and the assay also showed no cross-reactivity with abrin toxin.

A versatile approach for the immobilization of amines via copper-free “click” reaction between azido self-assembled monolayer and alkynyl Fischer carbene complex. Application to the detection of staphylococcal enterotoxin A antibody

A copper-free “click” reaction between an azido-terminated self-assembled monolayer (SAM) chemisorbed on planar gold-coated glass sensors and an alkynyl Fischer carbene complex yielded functionalized surfaces onto which facile and swift grafting of amine-containing molecules was achieved via aminolysis of the Fischer carbene moieties. The course of this process was conveniently monitored by Polarization-Modulation Infrared Reflection-Absorption Spectroscopy (PM-IRRAS), contact angle measurements and X-ray photoelectron spectroscopy (XPS). A study of the parameters involved in the covalent grafting of the model protein bovine serum albumin (BSA) to the Fischer carbene modified SAM was carried out. As an application, the bacterial toxin staphylococcal enterotoxin A (SEA) produced by some pathogenic strains of Staphylococcus aureus was immobilized on the gold chips and immunocapture of a polyclonal antibody raised against SEA was detected by PM-IRRAS so as to ultimately construct an optical immunosensor for the detection of this toxin in food samples.

Detection of cholera toxin in seafood using a ganglioside-liposome immunoassay

Microbiological contamination of foods continues to be a major concern in public health. Biological toxins are one class of important contaminants that can cause various human diseases. Outbreaks related to contamination by biological toxins or toxin-producing microorganisms have made it extremely important to develop rapid (approximately 20min), sensitive and cost-effective analytical methods. This paper describes the development of a sensitive bioassay for the detection of cholera toxin (CT) in selected seafood samples, using ganglioside-incorporated liposomes. In this study, the assays were run with food samples spiked with various concentrations of CT. The limit of detection (LOD) increased by a factor of about 10-20 in most food samples, compared with the LOD in the buffer system previously reported. However, the LOD of toxins in food samples (8 × 10-3 × 10(3)fg/mL for CT) was still comparable to, or lower than, that previously reported for other assays. The results from this study demonstrate that the bioassays using ganglioside-liposomes can detect the toxin directly in the field screening of food samples rapidly, simply and reliably, without the need for complex instrumentation.

Analytical chromatography for recovery of small amounts of staphylococcal enterotoxins from food

Sample preparation is an important element in the detection of toxins in food samples. In this work, a simple analytical sample preparation method for recovery of small amount of staphylococcal enterotoxin B (SEB) and staphylococcal enterotoxin A (SEA) in food samples was developed. Cation exchanger carboxymethylcellulose (CM) was used for small-scale batch chromatography isolation of SEB from infant formula and from mushrooms spiked with SEB. The resulting materials were analyzed for SEB by Western immunoblotting. Nearly all of the extraneous substances in the sample were removed by this procedure with no significant loss of the toxin. Using this method, even small amounts of SE (0.75 ng/g) can be recovered and immunologically analyzed by Western blotting or by ELISA with a very low background. Because this method is effective, rapid, simple and inexpensive, it has the potential to be a general method for the preparation of samples used for analysis of SEs.

Multitoxin biosensor–mass spectrometry analysis: a new approach for rapid, real-time, sensitive analysis of staphylococcal toxins in food

Biomolecular interaction analysis mass spectrometry (BIA–MS) was applied to detection of bacterial toxins in food samples. This two-step approach utilizes surface plasmon resonance (SPR) to detect the binding of the toxin(s) to antibodies immobilized on a surface of a sensor chip. SPR detection is then followed by identification of the bound toxin(s) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Staphylococcal enterotoxin B (SEB) was readily detected in milk and mushroom samples at levels of 1 ng/ml. In addition, non-specific binding of food components to the immobilized antibody and to the sensor chip surface was detected. To evaluate the applicability of BIA–MS in the analysis of materials containing multiple toxic components, sample containing both SEB and toxic-shock syndrome toxin-1 was analyzed. Both toxins were successfully and simultaneously detected through the utilization of multiaffinity sensor chip surfaces.