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Bisphenol F and aniline-based benzoxazine monomers were selected to fabricate basalt, glass and carbon fiber reinforced polybenzoxazine via vacuum infusion, respectively. The impacts of the type of fiber reinforcement on the resulting material properties of the fiber reinforced polymers (FRPs) were studied. FRPs exhibited a homogenous morphology with completely impregnated fibers and near-zero porosity. Carbon fiber reinforced polybenzoxazine showed the highest specific mechanical properties because of its low density and high modulus and strength. However, regarding the flammability, fire, smoke and toxicity properties, glass and basalt reinforced polybenzoxazine outperformed carbon fiber reinforced polybenzoxazine. This work offers a deeper understanding of how different types of fiber reinforcement affect polybenzoxazine-based FRPs and provides access to FRPs with inherently good fire, smoke and toxicity performance without the need for further flame retardant additives.
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The ubiquitous nitrite is a major analyte in the management of human health and environmental risks. The current analytical methods are complex techniques that do not fulfil the need for simple, robust and low-cost tools for on-site monitoring. Electrochemical reductase-based biosensors are presented as a powerful alternative, due to their good analytical performance and miniaturization potential. However, their real-world application is limited by the need of anoxic working conditions, and the standard oxygen removal strategies are incompatible with point-of-care measurements. Instead, a bienzymatic oxygen scavenger system comprising glucose oxidase and catalase can be used to promote anoxic conditions in aired environments. Herein, carbon screen-printed electrodes were modified with cytochrome c nitrite reductase together with glucose oxidase and catalase, so that nitrite cathodic detection could be performed by cyclic voltammetry under ambient air. The resulting biosensor displayed good linear response to the analyte (2-200 µM, sensitivity of 326 ± 5 mA M-1 cm-2 at -0.8 V; 0.8-150 µM, sensitivity of 511 ± 11 mA M-1 cm-2 at -0.5 V), while being free from oxygen interference and stable up to 1 month. Furthermore, the biosensor's catalytic response was unaffected by the presence of cyanide, a well-known inhibitor of heme-enzymes.
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In this paper we aim to demonstrate, as a proof-of-concept, the feasibility of the mass production of effective point of care tests for nitrite quantification in environmental, food and clinical samples. Following our previous work on the development of third generation electrochemical biosensors based on the ammonia forming nitrite reductase (ccNiR), herein we reduced the size of the electrodes' system to a miniaturized format, solved the problem of oxygen interference and performed simple quantification assays in real samples. In particular, carbon paste screen printed electrodes (SPE) were coated with a ccNiR/carbon ink composite homogenized in organic solvents and cured at low temperatures. The biocompatibility of these chemical and thermal treatments was evaluated by cyclic voltammetry showing that the catalytic performance was higher with the combination acetone and a 40°C curing temperature. The successful incorporation of the protein in the carbon ink/solvent composite, while remaining catalytically competent, attests for ccNiR's robustness and suitability for application in screen printed based biosensors. Because the direct electrochemical reduction of molecular oxygen occurs when electroanalytical measurements are performed at the negative potentials required to activate ccNiR (ca.-0.4V vs Ag/AgCl), an oxygen scavenging system based on the coupling of glucose oxidase and catalase activities was successfully used. This enabled the quantification of nitrite in different samples (milk, water, plasma and urine) in a straightforward way and with small error (1-6%). The sensitivity of the biosensor towards nitrite reduction under optimized conditions was 0.55 A M(-1) cm(-2) with a linear response range 0.7-370 µM.
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Citocromos a1/química , Citocromos c1/química , Grafito/química , Tinta , Nitrato Reductasas/química , Nitritos/análisis , Acetona/química , Animales , Técnicas Biosensibles , Butanonas/química , Catalasa/química , Catálisis , Agua Potable/análisis , Electrodos , Enzimas Inmovilizadas/química , Glucosa/química , Glucosa Oxidasa/química , Leche/química , Miniaturización , Nitritos/sangre , Nitritos/orina , Oxígeno/química , Pruebas en el Punto de Atención , Solventes/químicaRESUMEN
This work describes the fabrication of a new lactate biosensor. The strategy is based on the use of a novel hybrid nanomaterial for amperometric biosensors i.e. platinum nanoparticles (PtNps) supported on graphitized carbon nanofibers (PtNps/GCNF) prepared by chemical reduction of the Pt precursor at GCNF surfaces. The biosensors were constructed by covalent immobilization of lactate oxidase (LOx) onto screen printed carbon electrodes (SPCEs) modified with PtNps (PtNps/GCNF-SPCEs) using polyethyleneimine (PEI) and glutaraldehyde (GA). Experimental variables concerning both the biosensor design and the detection process were investigated for an optimal analytical performance. Lactate biosensors show good reproducibility (RSD 4.9%, n=10) and sensitivity (41,302±546) µA/Mcm(2), with a good limit of detection (6.9µM). Covalent immobilization of the enzyme allows the reuse of the biosensor for several measurements, converting them in a cheap alternative to the solid electrodes. The long-term stability of the biosensors was also evaluated. 90% of the signal was kept after 3months of storage at room temperature (RT), while 95% was retained after 18months at -20°C. These results demonstrate that the method provides sensitive electrochemical lactate biosensors where the stability of the enzymatic activity can be preserved for a long period of time in adequate storage conditions.
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Bebidas/análisis , Técnicas Biosensibles/métodos , Ácido Láctico/análisis , Nanofibras/química , Vino/análisis , Técnicas Biosensibles/instrumentación , Carbono/química , Electrodos , Enzimas Inmovilizadas/química , Diseño de Equipo , Análisis de los Alimentos/instrumentación , Análisis de los Alimentos/métodos , Grafito/química , Oxigenasas de Función Mixta/química , Oxigenasas de Función Mixta/metabolismo , Platino (Metal)/química , Reproducibilidad de los Resultados , TemperaturaRESUMEN
A novel biosensor for lactate has been developed, using screen-printed carbon electrodes (SPCE) and lactate oxidase (LOx). The active surface of the electrodes was modified using a dispersion of platinum nanoparticle decorated carbon nanofibers (PtNp-CNF) in poly(diallyldimethylammonium) chloride (PDDA) solution. In this way, sensitive, disposable, low cost and reliable hydrogen peroxide sensors were obtained. The immobilisation of LOx on top of these PtNp-CNF-PDDA/SPCEs resulted in amperometric biosensors with high operational stability. The sensitivity of the optimised lactate biosensor was 36.8 (mA/Mcm(2)) with a linear range of 25-1500 µM. The limit of detection was 11 µM (S/N=3). Reproducibility, selectivity and storage stability were also evaluated. Additionally, the stability of the biosensor was also predicted by a model based on thermal degradation. Finally, lactate in sweat and blood samples was determined in a sport test using LOx/PtNp-CNF-PDDA/SPCEs and commercial biosensors respectively. Based on these data, the validity of the sweat lactate for the determination of the lactate threshold is discussed.
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Enzimas Inmovilizadas/metabolismo , Ácido Láctico/análisis , Ácido Láctico/sangre , Oxigenasas de Función Mixta/metabolismo , Nanofibras/química , Polietilenos/química , Compuestos de Amonio Cuaternario/química , Técnicas Biosensibles/métodos , Carbono/química , Enzimas Inmovilizadas/química , Humanos , Ácido Láctico/metabolismo , Límite de Detección , Oxigenasas de Función Mixta/química , Nanofibras/ultraestructura , Pediococcus/enzimología , Platino (Metal)/química , Reproducibilidad de los Resultados , Sudor/químicaRESUMEN
The development of sensors to detect specific weak biological interactions is still today a challenging topic. Characteristics of carbohydrate-protein (lectin) interactions include high specificity and low affinity. This work describes the development of nanostructured impedimetric sensors for the detection of concanavalin A (Con A) binding to immobilized thiolated carbohydrate derivatives (D-mannose or D-glucose) onto screen-printed carbon electrodes (SPCEs) modified with gold nanoparticles. Thiolated D-galactose derivative was employed as negative control to evaluate the selectivity of the proposed methodology. After binding the thiolated carbohydrate to the nanostructured SPCEs, different functionalized thiols were employed to form mixed self-assembled monolayers (SAM). Electrochemical impedance spectroscopy (EIS) was employed as a technique to evaluate the binding of Con A to selected carbohydrates through the increase of electron transfer resistance of the ferri/ferrocyanide redox probe at the differently SAM modified electrodes. Different variables of the assay protocol were studied in order to optimize the sensor performance. Selective Con A determinations were only achieved by the formation of mixed SAMs with adequate functionalized thiols. Important differences were obtained depending on the chain lengths and functional groups of these thiols. For the 3-mercapto-1-propanesulfonate mixed SAMs, the electron transfer resistance varied linearly with the Con A concentration in the 2.2-40.0 µg mL(-1) range for D-mannose and D-glucose modified sensors. Low detection limits (0.099 and 0.078 pmol) and good reproducibility (6.9 and 6.1%, n=10) were obtained for the D-glucose and D-mannose modified sensors, respectively, without any amplification strategy.
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Técnicas Biosensibles , Concanavalina A/análisis , Glucosa/química , Oro/química , Manosa/química , Nanopartículas del Metal/química , Carbono/química , Impedancia Eléctrica , Electroquímica , ElectrodosRESUMEN
In this article, a disposable magnetic DNA sensor using an enzymatic amplification strategy for the detection of specific hybridization processes, based on the coupling of streptavidin-peroxidase to biotinylated target sequences, has been developed. A thiolated 19-mer capture probe was attached to gold coated ferric oxide nanoparticles and hybridization with the biotinylated target was allowed to proceed. Then, a streptavidin-peroxide was attached to the biotinylated target and the resulting modified gold coated ferric oxide nanoparticles were captured by a magnetic field on the surface of a home-made carbon screen printed electrode (SPE). Using hydroquinone as a mediator, a square wave voltammetric procedure was chosen to detect the hybridization process after the addition of hydrogen peroxide. Different aspects concerning the assay protocol and nanoparticles fabrication were optimized in order to improve the sensitivity of the developed methodology. A low detection limit (31 pM) with good stability (RSD=7.04%, n=10) was obtained without the need of polymerase chain reaction (PCR) amplification.
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Técnicas Biosensibles/instrumentación , Mapeo Cromosómico/instrumentación , Materiales Biocompatibles Revestidos/química , Compuestos Férricos/química , Hibridación in Situ/instrumentación , Magnetismo/instrumentación , Nanopartículas/química , Equipos Desechables , Diseño de Equipo , Análisis de Falla de Equipo , Oro/química , Nanopartículas/ultraestructura , Nanotecnología/instrumentaciónRESUMEN
This work describes the use of a PEDOT:PSS-based conductive polymer for designing AChE-based biosensors. The transducers were obtained directly by screen-printing a PEDOT:PSS suspension on the surface of thick film carbon electrodes. The obtained working electrodes showed a high conductivity when compared with electrodes modified with conventional mediators like cobalt phthalocyanine or tetracyanoquinodimethane. The PEDOT:PSS polymer was shown to be suitable for thiocholine oxidation, allowing the measurement of AChE activity at 100 mV vs Ag/AgCl. The high conductivity of PEDOT:PSS allowed the accurate detection of the organophosphate insecticide chlorpyrifos-oxon at concentrations as low as 4x10(-9)M, corresponding to an inhibition ratio of 5%.