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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.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , COVID-19 , Anticorpos , Aptâmeros de Nucleotídeos/química , Solventes Eutéticos Profundos , Glutens , Humanos , Limite de Detecção , Solventes/químicaRESUMO
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.
Assuntos
Lactoglobulinas , Hipersensibilidade a Leite , Alérgenos , Animais , Caseínas , Bovinos , Feminino , Leite , Hipersensibilidade a Leite/diagnósticoRESUMO
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.
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Técnicas Biossensoriais , Catálise , Eletroquímica , Ligação de Hidrogênio , SolventesRESUMO
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.
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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.
Assuntos
Aptâmeros de Nucleotídeos/química , Glutens/análise , Técnica de Seleção de Aptâmeros/métodos , Solventes/química , Aptâmeros de Nucleotídeos/metabolismo , Sequência de Bases , Biotinilação , Glutens/metabolismo , Água/químicaRESUMO
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.
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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.
Assuntos
Gorduras Insaturadas na Dieta/análise , Técnicas Eletroquímicas/instrumentação , Papel , Técnicas Eletroquímicas/métodos , Eletrodos , Desenho de Equipamento , Fenóis/análise , Reprodutibilidade dos TestesRESUMO
This paper describes for the first time the fabrication of pencil drawn electrodes (PDE) on paper platforms for capacitively coupled contactless conductivity detection (C(4) D) on electrophoresis microchips. PDE-C(4) D devices were attached on PMMA electrophoresis chips and used for detection of K(+) and Na(+) in human tear samples. PDE-C(4) D devices were produced on office paper and chromatographic paper platforms and their performance were thoroughly investigated using a model mixture containing K(+) , Na(+) , and Li(+) . In comparison with chromatographic paper, PDE-C(4) D fabricated on office paper has exhibited better performance due to its higher electrical conductivity. Furthermore, the detector response was similar to that recorded with electrodes prepared with copper adhesive tape. The fabrication of PDE-C(4) D on office paper has offered great advantages including extremely low cost (< $ 0.004 per unit), reduced fabrication time (< 5 min), and minimal instrumentation (pencil and paper). The proposed electrodes demonstrated excellent analytical performance with good reproducibility. For an inter-PDE comparison (n = 7), the RSD values for migration time, peak area, and separation efficiency were lower than 2.5, 10.5, and 14%, respectively. The LOD's achieved for K(+) , Na(+) , and Li(+) were 4.9, 6.8, and 9.0 µM, respectively. The clinical feasibility of the proposed approach was successfully demonstrated with the quantitative analysis of K(+) and Na(+) in tear samples. The concentration levels found for K(+) and Na(+) were, respectively, 20.8 ± 0.1 mM and 101.2 ± 0.1 mM for sample #1, and 20.4 ± 0.1 mM and 111.4 ± 0.1 mM for sample #2.
Assuntos
Cátions/análise , Eletroforese em Microchip/instrumentação , Eletroforese em Microchip/métodos , Papel , Lágrimas/química , Condutividade Elétrica , Eletrodos , Desenho de Equipamento , HumanosRESUMO
The hyphenation of electrochemical methods and optical methods in a single portable device is expected to be a challenging combination to enhance the information which can be gained on complex chemical systems. In this paper, a low-cost spectrophotometric device based on low-cost electronics integrated with an electroanalytical cell equipped with a screen printed electrode (SPE) and assembled exploiting a DIY approach, is presented. This easy to use device allowed spectrophotometric and electroanalytical measurements to be performed simultaneously providing simultaneous information and enabling concomitant comparison and autovalidation of the results collected. The analytical robustness and precision of the proposed system was successfully tested on solutions containing mixtures of Patent Blue (E-131) and Brilliant Blue (Erioglaucine E-133), two food dyes displaying optical and redox properties very similar to each other.
RESUMO
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.
Assuntos
Qualidade dos Alimentos , Líquidos Iônicos/química , Odorantes/análise , Técnicas de Microbalança de Cristal de Quartzo/métodos , Alquilação , Cinnamomum zeylanicum/química , Análise de Componente Principal , Temperatura , Compostos Orgânicos Voláteis/análise , Compostos Orgânicos Voláteis/químicaRESUMO
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.
Assuntos
Técnicas Eletroquímicas/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Ácido Ascórbico/isolamento & purificação , Compostos Azo/isolamento & purificação , Eletrodos , Desenho de Equipamento , Papel , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
This communication describes a simple and cost-effective method for fabricating glass microchannels by wet chemical etching using masks made by xurography in vinyl adhesive films. Analytical performance of microfluidic devices fabricated using the new approach was evaluated by microchip electrophoresis coupled to capacitively coupled contactless conductivity detection (C(4)D) and laser-induced fluorescence (LIF) detection.
Assuntos
Eletroforese em Microchip/instrumentação , Vidro/química , Microtecnologia/métodos , Adesivos/química , Desenho de EquipamentoRESUMO
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.
Assuntos
Técnicas Eletroquímicas/instrumentação , Gases/análise , Líquidos Iônicos/química , Fenol/análise , Compostos de Sulfidrila/análise , Ácidos/análise , Condutividade Elétrica , Técnicas Eletroquímicas/economia , Eletrodos , Desenho de Equipamento , Limite de Detecção , Papel , Temperatura , VolatilizaçãoRESUMO
The need for providing rapid and, possibly, on-the-spot analytical results in the case of intoxication has prompted researchers to develop rapid, sensitive, and cost-effective methods and analytical devices suitable for use in nonspecialized laboratories and at the point of need (PON). In recent years, the technology of paper-based microfluidic analytical devices (µPADs) has undergone rapid development and now provides a feasible, low-cost alternative to traditional rapid tests for detecting harmful compounds. In fact, µPADs have been developed to detect toxic molecules (arsenic, cyanide, ethanol, and nitrite), drugs, and drugs of abuse (benzodiazepines, cathinones, cocaine, fentanyl, ketamine, MDMA, morphine, synthetic cannabinoids, tetrahydrocannabinol, and xylazine), and also psychoactive substances used for drug-facilitated crimes (flunitrazepam, gamma-hydroxybutyric acid (GHB), ketamine, metamizole, midazolam, and scopolamine). The present report critically evaluates the recent developments in paper-based devices, particularly in detection methods, and how these new analytical tools have been tested in forensic and clinical toxicology, also including future perspectives on their application, such as multisensing paper-based devices, microfluidic paper-based separation, and wearable paper-based sensors.
Assuntos
Cocaína , Ketamina , Microfluídica , Toxicologia Forense , Dispositivos Lab-On-A-ChipRESUMO
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.
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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.
Assuntos
Aminoácidos/isolamento & purificação , Aminas Biogênicas/isolamento & purificação , Eletrodos , Eletroforese em Microchip/métodos , Aminoácidos/química , Cerveja/análise , Aminas Biogênicas/química , Cianetos/química , Estabilidade de Medicamentos , Concentração de Íons de Hidrogênio , Oxirredução , Compostos de Rutênio/químicaRESUMO
The recent developments on fieldable miniature mass spectrometers require efforts to produce easy-to-use and portable alternative tools to assist in point-of-care analysis. In this paper, the reagent-pencil (RP) technology, which has been used for solvent-free deposition of reagents in paper-based microfluidics, was combined with paper spray ionization mass spectrometry (PS-MS). In this approach, named RP-PS-MS, the PS triangular piece of paper was written with the reagent pencil, consisting of mixtures of graphite and bentonite (used as a support) and a reactive compound, and allowed to react with a given analyte from a sample matrix selectively. We conducted typical applications as proof-of-principles to verify the methodology's general usefulness in detecting small organic molecules in distinct samples. Hence, various aldehydes (2-furaldehyde, valeraldehyde, and benzaldehyde) in spiked cachaça samples (an alcoholic drink produced from fermentation/distillation of sugarcane juice) were promptly detected using a reagent pencil doped with 4-aminophenol (the reactive compound). Similarly, we recognized typical ginsenosides and triacylglycerols (TAGs) in ginseng aqueous infusions and soybean oil samples, respectively, using lithium chloride as the reactive compound. The results indicate that the reagent-pencil methodology is compatible with PS-MS and provides an easy and fast way to detect target analytes in complex samples. The advantage over the usual solution-based deposition of reagents lies in the lack of preparation or carrying different specific solutions for special applications, which can simplify operation, especially in point-of-care analysis with fieldable mass spectrometers.
RESUMO
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.
Assuntos
Eletroforese em Microchip/instrumentação , Catecóis/isolamento & purificação , Dopamina/isolamento & purificação , Eletroquímica , Eletroforese em Microchip/economia , Eletroforese em Microchip/métodos , Desenho de Equipamento , Reprodutibilidade dos Testes , Fatores de TempoRESUMO
Enzyme-linked immunosorbent assays are currently the most popular methods to quantify gluten in foods. Unfortunately, the antibodies used as specific receptors in such methods are not compatible with the usual solvents for the extraction of gluten proteins. In consequence, commercial tests require a high dilution of the sample after the extraction, increasing the limit of quantification and decreasing convenience. In this work, we have rationally truncated an aptamer capable of recognizing gliadin in a deep eutectic solvent (DES). The truncated aptamer is a 19-nucleotides-long DNA that minimizes self-hybridization, allowing the development of an electrochemical sandwich-based sensor for the quantification of gluten in the DES ethaline. The sensor incorporates two identical biotin-labeled truncated aptamers, one of which is immobilized on a carbon screen-printed electrode and the other reports the binding of gliadin after incubation in streptavidin-peroxidase. This sensor can detect gliadin in DES, with a dynamic range between 1 and 100 µg/L and an intra-assay coefficient of variation of 11%. This analytical performance allows the quantification of 20 µg of gluten/kg of food when 1 g of food is extracted with 10 mL of ethaline. We demonstrate the ability of this method to achieve the measurement of gluten in food samples, after the extraction with pure ethaline. The assay is useful for the analysis of residual gluten levels in foods, thus facilitating the evaluation of any potential health risk associated with the consumption of such food by people with celiac disease or other gluten-related disorders.