Application of Highly Sensitive Immunosensor Based on Optical Waveguide Light-Mode Spectroscopy (OWLS) Technique for the Detection of the Herbicide Active Ingredient Glyphosate.

The herbicide active ingredient glyphosate is the most widely applied herbicidal substance worldwide. Currently it is the market-leading pesticide, and its use is projected to further grow 4.5-fold between 2022 and 2029. Today, glyphosate use exceeds one megaton per year worldwide, which represents a serious environmental burden. A factor in the overall boost in the global use of glyphosate has been the spread of glyphosate-tolerant genetically modified (GM) crops that allow post-emergence applications of the herbicide on these transgenic crops. In turn, cultivation of glyphosate-tolerant GM crops represented 56% of the glyphosate use in 2019. Due to its extremely high application rate, xenobiotic behaviour and a water solubility (11.6 mg/mL at 25 °C) unusually high among pesticide active ingredients, glyphosate has become a ubiquitous water pollutant and a primary drinking water contaminant worldwide, presenting a threat to water quality. The goal of our research was to develop a rapid and sensitive method for detecting this herbicide active ingredient. For this purpose, we applied the novel analytical biosensor technique optical waveguide light-mode spectroscopy (OWLS) to the label-free detection of glyphosate in a competitive immunoassay format using glyphosate-specific polyclonal antibodies. After immobilising the antigen conjugate in the form of a glyphosate conjugated to human serum albumin for indirect measurement, the sensor chip was used in a flow-injection analyser system. For the measurements, an antibody stock solution was diluted to 2.5 µg/mL. During the measurement, standard solutions were mixed with the appropriate concentration of antibodies and incubated for 1 min before injection. The linear detection range and the EC50 value of the competitive detection method were between 0.01 and 100 ng/mL and 0.60 ng/mL, respectively. After investigating the indirect method, we tested the cross-reactivity of the antibody with glyphosate and structurally related compounds.

Current Trends in Mycotoxin Detection with Various Types of Biosensors.

One of the most important tasks in food safety is to properly manage the investigation of mycotoxin contamination in agricultural products and foods made from them, as well as to prevent its occurrence. Monitoring requires a wide range of analytical methods, from expensive analytical procedures with high-tech instrumentation to significantly cheaper biosensor developments or even single-use assays suitable for on-site monitoring. This review provides a summary of the development directions over approximately a decade and a half, grouped according to the biologically sensitive components used. We provide an overview of the use of antibodies, molecularly imprinted polymers, and aptamers, as well as the diversity of biosensors and their applications within the food industry. We also mention the possibility of determining multiple toxins side by side, which would significantly reduce the time required for the analyses.

Application of Electrochemical Biosensors for Determination of Food Spoilage.

Food security is significantly affected by the mass production of agricultural produce and goods, the growing number of imported foods, and new eating and consumption habits. These changed circumstances bring food safety issues arising from food spoilage to the fore, making food safety control essential. Simple and fast screening methods have been developed to detect pathogens and biomarkers indicating the freshness of food for safety. In addition to the traditional, sequential, chemical analytical and microbiological methods, fast, highly sensitive, automated methods suitable for serial tests have appeared. At the same time, biosensor research is also developing dynamically worldwide, both in terms of the analytes to be determined and the technical toolkit. Consequently, the rapid development of biosensors, including electrochemical-based biosensors, has led to significant advantages in the quantitative detection and screening of food contaminants. These techniques show great specificity for the biomarkers tested and provide adequate analytical accuracy even in complex food matrices. In our review article, we summarize, in separate chapters, the electrochemical biosensors developed for the most important food groups and the food safety issues they can ensure, with particular respect to meat and fish products, milk and dairy products, as well as alcoholic and non-alcoholic beverages.

Conformal Temperature/Impedance Sensing Patch Based on Graphene Materials for Nondestructive Detection of Fish Freshness.

Rapid nondestructive detection of fish freshness is essential to ensure food safety and nutrition. In this study, we demonstrate a conformal temperature/impedance sensing patch for temperature monitoring, as well as freshness classification during fish storage. The optimization of the flexible laser-induced graphene electrodes is studied based on both simulation and experimental validation, and dimensional accuracy of 5‰ and high impedance reproducibility are obtained. A laser-assisted thermal reduction technology is innovatively introduced to directly form a reduced graphene oxide-based temperature-sensitive layer on the surface of a flexible substrate. The comprehensive performance is superior to that of most reported temperature-sensitive devices based on graphene materials. As an application demonstration, the fabricated flexible dual-parameter sensing patch is conformed to the surface of a refrigerated fish. The patch demonstrates the ability to accurately sense low temperatures in a continuous 120 min monitoring, accompanied by no interference from high humidity. Meanwhile, the collected impedance data are imported into the support vector machine model to obtain a freshness classification accuracy of 93.07%. The conformal patch integrated with crosstalk-free dual functions costs less than $1 and supports free customization, providing a feasible methodology for rapid nondestructive detection or monitoring of food quality.

Hysteresis Dynamic Modeling and Analysis of Flexible Nano Silver-Polyvinyl Alcohol Humidity Sensor Based on the Microscopic Process and Langmuir-Fick Theory.

In recent years, advances in materials science and manufacturing technologies have facilitated the development of flexible sensors. However, there are still performance gaps between emerging flexible sensors and traditional silicon-based rigid sensors, especially lacking dynamic modeling and optimization analysis for addressing above challenges. This paper describes a hysteresis dynamic modeling method for flexible humidity sensors. Through inkjet printing and coating methods, the polyvinyl alcohol (PVA) sensitive layer and nano silver interdigital electrode are fabricated on flexible polyethylene naphthalate substrates. The performance characterization results show that the sensitivity and maximum hysteresis within the range of 12-98% relative humidity (RH) are -0.02167 MΩ/% RH and 2.7% RH, respectively. The sensor also has outstanding dynamic response ability and stability in a wide range of humidity variation. The hysteresis mechanism of flexible humidity sensors is theoretically analyzed from microscopic hysteresis processes, Langmuir monomolecular adsorption dynamic modeling, and Fick diffusion dynamic modeling. These hysteresis models provide a path for the hysteresis optimization of flexible PVA humidity sensors. Further exploration of the diffusion rate of water molecules and the proportion of PVA in ink represents promising hysteresis optimization directions of flexible humidity sensors based on PVA-sensitive material.

Development of an Immunofluorescent Capillary Sensor for the Detection of Zearalenone Mycotoxin.

A capillary-based immunofluorescence sensor was developed and incorporated in a flow injection analysis system. The light-guiding capillary was illuminated axially by a 473 nm/5 mW solid state laser through a tailored optofluidic connector. High sensitivity of the system was achieved by efficiently collecting and detecting the non-guided fluorescence signal scattered out along the wall of the capillary. The excitation was highly suppressed with bandpass and dichroic filters by simultaneously exploiting the guiding effect inside the capillary. The glass capillary used as a measuring cell was silanized in liquid phase by 3-aminopropyltriethoxysilane (APTS), and the biomolecules were immobilized using glutaraldehyde inside the capillary. The applicability of the developed system was tested with a bovine serum albumin (BSA)-anti-BSA-IgG model-molecule pair, using a fluorescently labeled secondary antibody. Based on the results of the BSA-anti-BSA experiments, a similar setup using a primary antibody specific for zearalenone (ZON) was established, and a competitive fluorescence measurement system was developed for quantitative determination of ZON. For the measurements, 20 µg/mL ZON-BSA conjugate was immobilized in the capillary, and a 1:2500 dilution of the primary antibody stock solution and a 2 µg/mL secondary antibody solution were set. The developed capillary-based immunosensor allowed a limit of detection (LOD) of 0.003 ng/mL and a limit of quantification (LOQ) of 0.007 ng/mL for ZON in the competitive immunosensor setup, with a dynamic detection range of 0.01-10 ng/mL ZON concentrations.

Biosensors for Deoxynivalenol and Zearalenone Determination in Feed Quality Control.

Mycotoxin contamination of cereals used for feed can cause intoxication, especially in farm animals; therefore, efficient analytical tools for the qualitative and quantitative analysis of toxic fungal metabolites in feed are required. Current trends in food/feed analysis are focusing on the application of biosensor technologies that offer fast and highly selective and sensitive detection with minimal sample treatment and reagents required. The article presents an overview of the recent progress of the development of biosensors for deoxynivalenol and zearalenone determination in cereals and feed. Novel biosensitive materials and highly sensitive detection methods applied for the sensors and the application of these sensors to food/feed products, the limit, and the time of detection are discussed.

Optical waveguide light-mode spectroscopy immunosensors for environmental monitoring.

Coupling the high specificity of the immunoanalytical reaction with the high sensitivity of optical waveguide light-mode spectroscopy (OWLS) detection gives the possibility to develop immunosensors with in most cases a definitely lower detection limit than traditionally used immunoassays. Measurements were performed on the sensitized surface of optical waveguide grating coupler sensors (2400 lines/mm grating). The OWLS technique is based on the precise measurement of the resonance angle of a polarized laser light (632.8 nm), diffracted by a grating and incoupled into a thin waveguide. The effective refractive index, determined from the resonance incoupling angle detected at high accuracy, allows determination of layer thickness and coverage (or mass) of the adsorbed or bound material with ultrahigh sensitivity. OWLS immunosensors were developed as label-free immunosensors with an amino group modified SiO(2)-TiO(2) sensor surface on which the immunoreactants could be anchored. One of the components of the antibody-antigen complex was chemically bound on the sensor surface, allowing noncompetitive or competitive detection of the analytes. To illustrate that the resulting immunosensors are suitable for the determination of small and large molecular weight analytes, OWLS sensor formats were applied for quantitative detection of a herbicide active ingredient trifluralin, a Fusarium mycotoxin zearalenone, and an egg yolk protein of key importance in endocrine regulation, vitellogenin.

Optical waveguide lightmode spectroscopy technique-based immunosensor development for aflatoxin B1 determination in spice paprika samples.

Optical waveguide lightmode spectroscopy (OWLS) technique has been applied to label-free detection of aflatoxin B1 in a competitive immunoassay format, with the aim to compare the analytical goodness of the developed OWLS immunosenor with HPLC and enzyme-linked immunosorbent assay (ELISA) methods for the detection of aflatoxin in spice paprika matrix. We have also assessed applicability of the QuEChERS method prior to ELISA measurements, and the results were compared to those obtained by traditional solvent extraction followed by immunoaffinity clean-up. The AFB1 content of sixty commercial spice paprika samples from different countries were measured with the developed and optimized OWLS immunosensor. Comparing the results from the indirect immunosensor to that obtained by HPLC or ELISA provided excellent correlation (with regression coefficients above 0.94) indicating that the competitive OWLS immunosensor has a potential for quick determination of aflatoxin B1 in paprika samples.