A smartphone aptasensor for fipronil detection in honey samples.

In this study, an electrochemical smartphone-based aptasensor for the determination of fipronil was developed by modifying a screen-printed carbon electrode (SPCE). Fipronil is a broad-spectrum insecticide that has been widely used in various applications such as agriculture, veterinary, and household pest control. Recently, its use has raised concerns over the potential impact on the environment and human health. The absence of effective methods for this purpose poses a significant obstacle. To tackle this problem, we have developed a cutting-edge aptamer-based portable sensor capable of rapidly and conveniently detecting fipronil in situ. Considering that the detection of small molecules, such as fipronil, can be a challenging task, a competitive replacement assay was set up based on the aptamer's preference for the free form of fipronil over the immobilized one on the electrode. The analytical performance provided by the sensor on standard solutions of a known fipronil content made it possible to estimate a limit of detection (LOD) equal to 1.07 µg kg-1 and a limit of quantification (LOQ) of 3.21 µg kg-1. Selectivity tests were conducted using atrazine as a possible interferent. The use and performance of the developed portable aptasensor was assessed on honey samples, which were simultaneously analyzed using an HPLC-MS method. This aptasensor could be an affordable and effective tool for accurately quantifying fipronil not only in honey samples but also in other food products.

Synergistic Applications of Graphene-Based Materials and Deep Eutectic Solvents in Sustainable Sensing: A Comprehensive Review.

The recently explored synergistic combination of graphene-based materials and deep eutectic solvents (DESs) is opening novel and effective avenues for developing sensing devices with optimized features. In more detail, remarkable potential in terms of simplicity, sustainability, and cost-effectiveness of this combination have been demonstrated for sensors, resulting in the creation of hybrid devices with enhanced signal-to-noise ratios, linearities, and selectivity. Therefore, this review aims to provide a comprehensive overview of the currently available scientific literature discussing investigations and applications of sensors that integrate graphene-based materials and deep eutectic solvents, with an outlook for the most promising developments of this approach.

Electrochemical Evaluation of Tyrosinase Enzymatic Activity in Deep Eutectic Solvent and Aqueous Deep Eutectic Solvent.

The use of green, inexpensive, and biodegradable deep eutectic solvents as nonaqueous solvents and electrolytes could be a useful way to potentially improve the enzyme biosensor performance as well as a profitable strategy to extend their use in the gas phase. However, enzyme activity in these media, although fundamental for their implementation in electrochemical analysis, is still almost unexplored. In this study, an electrochemical approach was employed to monitor tyrosinase enzyme activity in a deep eutectic solvent. This study was performed in a DES consisting of choline chloride (ChCl) as a hydrogen bond acceptor (HBA) and glycerol as a hydrogen bond donor (HBD), while phenol was chosen as the prototype analyte. The tyrosinase enzyme was immobilized on a gold-nanoparticle-modified screen-printed carbon electrode, and its activity was monitored following the reduction current of orthoquinone produced by the tyrosinase biocatalysis of phenol. This work represents a first step toward the realization of green electrochemical biosensors capable of operating in both nonaqueous and gaseous media for the chemical analysis of phenols.

A portable electrochemiluminescence aptasensor for ß-lactoglobulin detection.

Cow's milk allergy is one of the most common food allergies in children with a prevalence of around 2.5%. Milk contains several allergens; the main ones are caseins and ß-lactoglobulin (ß-LG). At regulatory level, ß-LG is not explicitly named, but milk is included in the list of substances or products causing allergies or intolerances. Hence, the presence of ß-LG can be a useful marker for determining the presence of milk in food. In this work, we present an aptasensor based on electrochemiluminescence (ECL) for the quantification of ß-LG in real food matrices displaying integrated advantages consisting of high specificity, good sensitivity, portability, and cost effectiveness. The performance and applicability of this sensor were tested by analyzing a sample of skimmed milk and an oat-based drink proposed as a vegetable substitute for milk of animal origin. We obtained a linear correlation between the intensity of the signal and the concentration of ß-LG standard solutions (y = x * 0.00653 + 1.038, R2 = 0.99). The limit of detection (LOD) and the limit of quantification (LOQ) were found to be 1.36 and 4.55 µg L-1, respectively.

Paper-based aptamer-antibody biosensor for gluten detection in a deep eutectic solvent (DES).

Paper has been widely employed as cheap material for the development of a great number of sensors such as pregnancy tests, strips to measure blood sugar, and COVID-19 rapid tests. The need for new low-cost analytical devices is growing, and consequently the use of these platforms will be extended to different assays, both for the final consumer and within laboratories. This work describes a paper-based electrochemical sensing platform that uses a paper disc conveniently modified with recognition molecules and a screen-printed carbon electrode (SPCE) to achieve the detection of gluten in a deep eutectic solvent (DES). This is the first method coupling a paper biosensor based on aptamers and antibodies with the DES ethaline. Ethaline proved to be an excellent extraction medium allowing the determination of very low gluten concentrations. The biosensor is appropriate for the determination of gluten with a limit of detection (LOD) of 0.2 mg L-1 of sample; it can detect gluten extracted in DES with a dynamic range between 0.2 and 20 mg L-1 and an intra-assay coefficient of 10.69%. This approach can be of great interest for highly gluten-sensitive people, who suffer from ingestion of gluten quantities well below the legal limit, which is 20 parts per million in foods labeled gluten-free and for which highly sensitive devices are essential.

Deep Eutectic Solvents (DESs) and Their Application in Biosensor Development.

Deep Eutectic Solvents (DESs) are a new class of solvents characterized by a remarkable decrease in melting point compared to those of the starting components. The eutectic mixtures can be simply prepared by mixing a Hydrogen Bond Acceptor (HBA) with a Hydrogen Bond Donor (HBD) at a temperature of about 80 °C. They have found applications in different research fields; for instance, they have been employed in organic synthesis, electrochemistry, and bio-catalysis, showing improved biodegradability and lower toxicity compared to other solvents. Herein, we review the use of DESs in biosensor development. We consider the emerging interest in different fields of this green class of solvents and the possibility of their use for the improvement of biosensor performance. We point out some promising examples of approaches for the assembly of biosensors exploiting their compelling characteristics. Furthermore, the extensive ability of DESs to solubilize a wide range of molecules provides the possibility to set up new devices, even for analytes that are usually insoluble and difficult to quantify.

Modified Screen Printed Electrode Suitable for Electrochemical Measurements in Gas Phase.

We describe a convenient assembly for screen printed carbon electrodes (SPCE) suitable for analyses in gaseous samples which are of course lacking in supporting electrolytes. It consists of a circular crown of filter paper, soaked in a RTIL or a DES, placed upon a disposable screen printed carbon cell, so as to contact the outer edge of the carbon disk working electrode, as well as peripheral counter and reference electrodes. The electrical contact between the paper crown soaked in RTIL or DES and SPCE electrodes is assured by a gasket, and all components are installed in a polylactic acid holder. As a result of this configuration, a sensitive, fast-responding, membrane-free gas sensor is achieved where the real working electrode surface is the boundary zone of the carbon working disk contacted by the paper crown soaked in the polyelectrolyte. This assembly provides a portable and disposable electrochemical platform, assembled by the easy immobilization onto a porous and inexpensive supporting material such as paper of RTILs or DESs which are characterized by profitable electrical conductivity and negligible vapor pressure. The electroanalytical performance of this device was evaluated by voltammetric and flow injection analyses of oxygen which was chosen as prototype of electroactive gaseous analytes. The results obtained pointed out that this assembly is very profitable for the analysis of gaseous atmospheres, especially when used as detector for FIA in gaseous streams.

Characterization of Inherently Chiral Electrosynthesized Oligomeric Films by Voltammetry and Scanning Electrochemical Microscopy (SECM).

A voltammetric and scanning electrochemical microscopy (SECM) investigation was performed on an inherently chiral oligomer-coated gold electrode to establish its general properties (i.e., conductivity and topography), as well as its ability to discriminate chiral electroactive probe molecules. The electroactive monomer (S)-2,2'-bis(2,2'-bithiophene-5-yl)-3,3'-bibenzothiophene ((S)-BT2T4) was employed as reagent to electrodeposit, by cyclic voltammetry, the inherently chiral oligomer film of (S)-BT2T4 (oligo-(S)-BT2T4) onto the Au electrode surface (resulting in oligo-(S)-BT2T4-Au). SECM measurements, performed in either feedback or competition mode, using the redox mediators [Fe(CN)6]4- and [Fe(CN)6]3- in aqueous solutions, and ferrocene (Fc), (S)-FcEA, (R)-FcEA and rac-FcEA (FcEA is N,N-dimethyl-1-ferrocenylethylamine) in CH3CN solutions, indicated that the oligomer film, as produced, was uncharged. The use of [Fe(CN)6]3- allowed establishing that the oligomer film behaved as a porous insulating membrane, presenting a rather rough surface. This was inferred from both the approach curves and linear and bidimensional SECM scans, which displayed negative feedback effects. The oligomer film acquired semiconducting or fully conducting properties when the Au electrode was biased at potential more positive than 0.6 V vs. Ag|AgCl|KCl. Under the latter conditions, the approach curves displayed positive feedback effects. SECM measurements, performed in competition mode, allowed verifying the discriminating ability of the oligo-(S)-BT2T4 film towards the (S)-FcEA and (R)-FcEA redox mediators, which confirmed the results obtained by cyclic voltammetry. SECM linear scans indicated that the enantiomeric discriminating ability of the oligo-(S)-BT2T4 was even across its entire surface.

A Portable Setup for the Voltammetric Determination of Total Mercury in Fish with Solid and Nanostructured Gold Electrodes.

A simple procedure for field fish sample pretreatment was developed. This treatment in combination with square wave anodic stripping voltammetry (SW-ASV) with solid gold electrodes (SGE) and gold nanoparticle-modified glassy carbon electrodes (AuNPs-GCE) was applied for the determination of total mercury content. A certified reference material (CRM, Tuna Fish BCR 463), ten freeze-dried samples of canned tuna and two fresh fish samples were analysed both with a bench-top voltammetric analyser after microwave digestion and with a portable potentiostat after mild eating using a small commercial food warmer. The results obtained by the two SW-ASV approaches and by a Direct Mercury Analyser (DMA), the official method for mercury determination, were in very good agreement. In particular, (i) the results obtained with in field procedure are consistent with those obtained with the conventional microwave digestion; (ii) the presence of gold nanoparticles on the active electrode surface permits an improvement of the analytical performance in comparison to the SGE: the Limit of Quantification (LOQ) for mercury in fish-matrix was 0.1 µg L-1 (Hg cell concentration), corresponding to 0.06 mg kg-1 wet fish, which is a performance comparable to that of DMA. The pretreatment proposed in this study is very easy and applicable to fresh fish; in combination with a portable potentiostat, it proved to be an interesting procedure for on-site mercury determination.

Selection of Anti-gluten DNA Aptamers in a Deep Eutectic Solvent.

Herein, we show the feasibility of using deep eutectic solvents as a faster way of selecting aptamers targeting poorly water-soluble species. This unexplored concept is illustrated for gluten proteins. In this way, aptamer-based gluten detection can be performed directly in the extraction media with improved detectability. We envision deep implications for applications not only in food safety control but also in biomedicine.

Digitally Controlled Procedure for Assembling Fully Drawn Paper-Based Electroanalytical Platforms.

A simple, reliable, and low-cost fabrication method is proposed here for assembling paper-based electrochemical devices (PEDs) using a commercial desktop digitally controlled plotter/cutter, together with ordinary writing tools. Permanent markers (tips of 1 mm) were used to create effective hydrophobic barriers on paper, while micromechanical pencils (mounting 4B graphite leads, 0.5 mm in diameter) were adopted for automatically drawn precise reference, counter, and working carbon electrodes. Fabrication parameters, such as writing pressure and speed, were first optimized, and the electrochemical performance of these devices was then evaluated by using potassium hexacyanoferrate(II) as redox probe. The good interdevice reproducibility (4.8%) displayed by the relevant voltammetric responses confirmed that this strategy can be profitably adopted to easily assemble paper-based electrochemical devices in a highly flexible manner. The simplicity of the instrumentation used and the low cost of each single device (about $0.04), together with the speed of fabrication (about 2 min), are other important features of the proposed strategy. Finally, to confirm the effectiveness of this prototyping method for the analysis of real samples and rapid controls, PEDs assembled by this simple approach were successfully exploited for the analysis of vitamin B6 in food supplements.

Oxidative behavior of (+)-catechin in the presence of inactive dry yeasts: a comparison with sulfur dioxide, ascorbic acid and glutathione.

BACKGROUND: The antioxidant capacity of an inactive dry yeast preparation (YD) was investigated by conventional analytical methods (spectrophotometry, high-performance liquid chromatography) as well as by cyclic voltammetry in a (+)-catechin model wine and compared with that of some of the most common antioxidants found in wine: sulfur dioxide, ascorbic acid and glutathione. RESULTS: Sulfur dioxide was the highest-performing substance in protecting (+)-catechin against browning, followed by ascorbic acid and the YD preparation. Sulfites were the only antioxidant whose activity was clearly detectable in the model wines after 29 days of storage. Voltammetric studies demonstrated that the antioxidant capacity of the products tested was connected to their intrinsic characteristics and their molar concentrations (catechin/antioxidant molar ratio). CONCLUSION: The YD preparation displayed a certain ability to protect polyphenols against browning. The antioxidant activity of YDs towards (+)-catechin appeared to be based on different mechanisms with respect to that of the other products tested: the insoluble portion of these preparations (cell wall residues) might have a non-negligible role, even if the ability of YDs to release compounds able to suppress oxidation cannot be rejected. Direct comparison of the different antioxidants led to interesting indications concerning their mechanism of action in wine-like solution, depending on their concentration and intrinsic characteristics. © 2017 Society of Chemical Industry.

Simple pencil-drawn paper-based devices for one-spot electrochemical detection of electroactive species in oil samples.

We propose here simple electrochemical cells assembled with electrodes pencil drawn on paper for conducting one-spot tests enabling olive oil to be easily distinguished from other vegetable oils. They consist of small circular pads of hydrophilic paper defined by hydrophobic barriers, these last printed by using custom-designed rubber stamps, where working, reference, and counterelectrodes are drawn by pencil leads. These cells were first wetted with a small volume of aqueous electrolyte, avoiding coating of the upper surface of electrodes. A controlled volume of edible oil samples was then applied on top of the moist cell. The results found proved that these devices can be adopted as effective platforms suitable for the detection of electroactive compounds present in edible oils. In fact, they allow voltammetric profiles to be recorded not only for the oxidation of water soluble species (ortho-diphenols, as well as some monophenols and polyphenols) present in olive oils, but also for electroactive hydrophobic components (e.g., α-tocopherol) present in sunflower oils, which were chosen as model of seed oils. The whole of collected findings pointed out that simple one-spot tests performed by these devices enable olive oils to be easily distinguished from other edible oils on the basis of their clearly different voltammetric profiles. A satisfactory interdevice reproducibility (±13%) was estimated by applying strictly similar extra virgin olive oil samples onto seven different cells carefully prepared by the same procedure. An operating mechanism able to account for the detection of also electroactive hydrophobic compounds present in oils is proposed.

Room temperature ionic liquids as useful overlayers for estimating food quality from their odor analysis by quartz crystal microbalance measurements.

An array of quartz crystals coated with different room-temperature ionic liquids (RTILs) is proposed for the analysis of flavors by quartz crystal microbalance (QCM) measurements. Seven RTILs were adopted as sensing layers, all containing imidazolium or phosphonium cations, differing from one another in the length and branching of alkyl groups and neutralized by different anions. The array was at first applied to the analysis of 31 volatile organic compounds (VOCs), such as alcohols, phenols, aldehydes, esters, ketones, acids, amines, hydrocarbons and terpenes, chosen as representative components of a wide variety of food flavors. Multivariate data analysis by the principal component analysis (PCA) approach of the set of the corresponding responses led to separated clusters for these different chemical categories. To further prove the good performance of the RTIL-coated quartz crystal array as an "electronic nose", it was applied to the analysis of headspaces from cinnamon samples belonging to different botanical varieties ( Cinnamon zeylanicum and Cinnamon cassia ). PCA applied to responses recorded on different stocks of samples of both varieties showed that they could be fully discriminated.

Pencil-drawn paper supported electrodes as simple electrochemical detectors for paper-based fluidic devices.

A simple procedure for preparing inexpensive paper-based three-electrode electrochemical cells is described here. They consist of small circular pads of hydrophilic paper defined by hydrophobic barriers printed on paper with wax-based ink. The back face of these pads is insulated by thermally laminating a polyethylene layer and working, reference and counter electrodes are drawn on paper by using commercial pencil leads. At last, a controlled volume of sample containing a supporting electrolyte was laid to soak in paper channels. Their performance was evaluated by assaying these devices as both simple cells suitable for recording voltammograms on static samples and low-cost detectors for flowing systems. Voltammetric tests, conducted by using potassium hexacyanoferrate(II) as model prototype, were also exploited for identifying the brand and softness of graphite sticks enabling paper to be marked with lines displaying the best conductivity. By taking advantage of the satisfactory information thus gained, pencil drawn electrodes were tested as amperometric detectors for the separation of ascorbic acid and sunset yellow, which were chosen as prototype electroactive analytes because they are frequently present concomitantly in several food matrices, such as soft drinks and fruit juices. This separation was performed by planar thin layer chromatography conducted on microfluidic paper-based devices prepared by patterning on filter paper two longitudinal hydrophobic barriers, once again printed with wax-based ink. Factors affecting both separation and electrochemical detection were examined and optimised, with best performance achieved by using a 20 mM acetate running buffer (pH 4.5) and by applying a detection potential of 0.9 V. Under these optimum conditions, the target analytes could be separated and detected within 6 min. The recorded peaks were well separated and characterized by good repeatability and fairly good sensitivity, thus proving that this approach is indeed suitable for rapidly assembling inexpensive and reliable electrochemical detectors for flow analysis systems.

Electrochemical gas sensors based on paper-supported room-temperature ionic liquids for improved analysis of acid vapours.

A prototype of a fast-response task-specific amperometric gas sensor based on paper-supported room-temperature ionic liquids (RTILs) is proposed here for improved analysis of volatile acid species. It consists of a small filter paper foil soaked with a RTIL mixture containing an ionic liquid whose anion (acetate) displays a basic character, upon which three electrodes are screen printed by carbon ink profiting from a suitable mask. It takes advantage of the high electrical conductivity and negligible vapour pressure of RTILs and of their easy immobilization into a porous and inexpensive supporting material such as paper. The performance of this device, used as a wall-jet amperometric detector for flow injection analyses of headspace samples in equilibrium with aqueous solutions at controlled concentrations, was evaluated for phenol and 1-butanethiol vapours which were adopted as model acid gaseous analytes. The results obtained showed that the quite high potentials required for the detection of these analytes are lowered significantly, thanks to the addition of the basic acetate RTIL. In such a way, overlap with the medium discharge is avoided, and the possible adverse effect of interfering species is minimised. The sensor performance was quite satisfactory (detection limits, ca. 0.3 µM; dynamic range, ca. 1-200 µM, both referred to solution concentrations; correlation coefficients in the range 0.993-0.997; repeatability, ± 6% RSD; long-term stability, 9%); thus suggesting the possible use of this device for manifold applications.

An Effective Label-Free Electrochemical Aptasensor Based on Gold Nanoparticles for Gluten Detection.

Nanomaterials can be used to modify electrodes and improve the conductivity and the performance of electrochemical sensors. Among various nanomaterials, gold-based nanostructures have been used as an anchoring platform for the functionalization of biosensor surfaces. One of the main advantages of using gold for the modification of electrodes is its great affinity for thiol-containing molecules, such as proteins, forming a strong Au-S bond. In this work, we present an impedimetric biosensor based on gold nanoparticles and a truncated aptamer for the quantification of gluten in hydrolyzed matrices such as beer and soy sauce. A good relationship between the Rct values and PWG-Gliadin concentration was found in the range between 0.1-1 mg L-1 of gliadin (corresponding to 0.2-2 mg L-1 of gluten) with a limit of detection of 0.05 mg L-1 of gliadin (corresponding to 0.1 mg L-1 of gluten). The label-free assay was also successfully applied for the determination of real food samples.

Kinetics of electron transfer reactions by humic substances: Implications for their biogeochemical roles and determination of their electron donating capacity.

Humic substances (HS) possess redox active groups covering a wide range of potentials and are used by facultative anaerobic microorganisms as electron acceptors. To serve as suitable electron shuttles for anaerobic respiration, HS should be able to re-oxidize relatively quickly to prevent polarization of the surrounding medium. Mediated electrochemical oxidation and decolorization assays, based on the reduction of the radical ion of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS•-) allow to determine the electron donating capacity (EDC) of HS, but uncertainties remain about the reaction time that should be allowed to obtain environmentally meaningful EDC values. In this work, we performed a kinetic analysis of the time trend of the reduction of ABTS•- by HS by Vis and Electron Paramagnetic Resonance (EPR) spectroscopies and by cyclic voltammetry. We found evidences of two concomitant separate mechanisms of electron exchange: a fast and a slow transfer processes which may have different environmental roles. These results can set a base to identify the appropriate conditions for the spectrophotometric determination of the fast and slow components of the EDC of HS.

A modified electrode for the electrochemical detection of biogenic amines and their amino acid precursors separated by microchip capillary electrophoresis.

The use of a mixed-valent ruthenium oxide/hexacyanoruthenate polymeric film electrochemically deposited onto glassy carbon electrodes is proposed here for the detection of biogenic amines and their amino acid precursors, following their separation by microchip capillary electrophoresis. The ability of this ruthenium coating to electrocatalyze the oxidation of aliphatic and heterocyclic amines, as well as their amino acid precursors, was checked by using ethanolamine, tryptamine and tryptophane as prototype compounds and adopting a 25 mM sulphuric acid as the electrolyte in the detection cell, where a constant potential of 1.05 V versus Ag/AgCl, 3 M KCl was applied to the modified working electrode. Optimization of parameters affecting both detection and separation steps led to satisfactory separations when performed by using a 20 mM phosphate running buffer (pH 2.5) and applying a high voltage of 2.5 kV both in the separation and in the electrokinetic injection (duration 4 s). The recorded peaks were characterized by good repeatability (RSD ≤ 3.6%), high sensitivity and a wide linear range. Detection limits of 23 µM (1.4 mg/L), 27 µM (4.3 mg/L) and 34 µM (6.8 mg/L) were inferred for ethanolamine, tryptamine and tryptophane, respectively. The approach proposed here was also applied for the analysis of some double malt dark beers spiked with a controlled amount of the analytes considered.

A simple approach to the hydrodynamic injection in microchip electrophoresis with electrochemical detection.

A simple hydrodynamic injection method is proposed here for microchip CE coupled to electrochemical detection. It is based on the use of a precise syringe pump to push the sample into the microfluidic circuit, accompanied by the application of a secondary electric field to the injection channel, soon after the end of the injection step. In such a way, any counter pressure effect taking place when the sample plug enters the micrometric channel is prevented. Suitable optimization of this secondary electric field enables pushing of sample excess to be avoided and a narrow sample plug during the separation step to be maintained. Best conditions for hydrodynamic injection were achieved injecting catechol as model analyte by pressure with a syringe pump set at a flow rate of 8 microL/min for 6 s and applying to the injection channel a secondary high voltage of 700 V soon after the injection was completed. The reliability of this injection procedure has been proved by comparing electropherograms found for samples containing either catechol alone or catechol and dopamine together with those recorded under the same conditions by electrokinetic injection. Repeatability, expressed as RSD and estimated for seven replicate injections, turned out to be 2.1% for peak height of catechol used as single analyte and 0.9 and 1.1% for catechol and dopamine respectively, simultaneously injected.