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Herein, we present the first 3D-printed electrochemical portable biodevice for the detection of monkeypox virus (MKPV). The electrochemical device consists of two biosensors: an immunosensor and a genosensor specifically designed for the detection of the protein A29 and a target DNA of MKPV, respectively. The electrodes were manufactured using lab-made ultraflexible conductive filaments composed of carbon black, recycled PLA from coffee pods, and castor oil as a plasticizer. The sensors created through 3D printing technology exhibited good reproducibility and repeatability of analytical responses. Furthermore, both the immunosensor and genosensor demonstrated excellent MKPV detection capabilities, with a linear range from 0.01 to 1.0 µmol L-1 for the antigen and 0.1 to 20.0 µmol L-1 for the DNA target. The biosensors achieved limits of detection of 2.7 and 29 nmol L-1 for the immunosensor and genosensor, respectively. Interference tests conducted with the biosensors demonstrated their selectivity for MKPV. Moreover, analyses of fortified human serum samples showed recoveries close to 100%, confirming the absence of significant matrix effects for MKPV analysis. Therefore, the 3D-printed multiplex device represents a viable and highly promising alternative for on-site, portable, and rapid point-of-care MKPV monitoring.
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This work emphasizes the utilization of biochar, a renewable material, as an interesting platform for anchoring redox mediators and bioreceptors in the development of economic, environmentally friendly biosensors. In this context, Fe(III) ions were preconcentrated on highly functionalized activated biochar, allowing the stable synthesis of Prussian blue nanostructures with an average size of 58.3 nm. The determination of glucose was carried out by indirectly monitoring the hydrogen peroxide generated through the enzymatic reaction, followed by its subsequent redox reaction with reduced Prussian blue (also known as Prussian white) in a typical electrochemical-chemical mechanism. The EDC/NHS (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-Hydroxysuccinimide) pair was employed for the stable covalent immobilization of the enzyme on biochar. The biosensor demonstrated good enzyme-substrate affinity, as evidenced by the Michaelis-Menten apparent kinetic constant (4.16 mmol L-1), and analytical performance with a wide linear dynamic response range (0.05-5.0 mmol L-1), low limits of detection (0.94 µmol L-1) and quantification (3.13 µmol L-1). Additionally, reliable repeatability, reproducibility, stability, and selectivity were obtained for the detection of glucose in both real and spiked human saliva and blood serum samples.
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Técnicas Biosensibles , Carbón Orgánico , Ferrocianuros , Glucosa , Nanoestructuras , Ferrocianuros/química , Técnicas Biosensibles/métodos , Nanoestructuras/química , Carbón Orgánico/química , Glucosa/análisis , Glucosa/química , Humanos , Enzimas Inmovilizadas/química , Glucosa Oxidasa/química , Glucosa Oxidasa/metabolismo , Glucemia/análisis , Peróxido de Hidrógeno/química , Peróxido de Hidrógeno/análisis , Límite de DetecciónRESUMEN
Platinum nanoparticles supported by carbon nanotubes were obtained by a simple chemical route and used for preparation of electrochemical sensor towards caffeine determination. Carbon nanotubes were used before and after an acid treatment, yielding two different materials. Morphological and structural characterization of these materials showed platinum nanoparticles (size around 12 nm) distributed randomly along carbon nanotubes. Modified electrodes were directly prepared through a dispersion of these materials. Voltammetric studies in the presence of caffeine revealed an electrocatalytic effect of platinum oxides, electrochemically produced from the chemical oxidation of the platinum nanoparticles. This behavior was explored in the development a selective method for caffeine determination based on platinum oxide reduction at a lower potential value (+0.45 V vs. Ag/AgCl). Using the best set of experimental conditions, it was shown a linear relationship for the caffeine concentration ranging from 5.0 to 25 µmol L-1 with a sensitivity of 449 nA L µmol-1. Limits of detection and quantification of 0.54 and 1.80 µmol L-1 were calculated, respectively. Recovery values for real samples of caffeine pharmaceutical formulations between 98.6% and 101.0% (n = 3) were obtained using the proposed procedure. Statistical calculations showed good concordance (95% confidence level) between the added and recovery values.
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Cafeína , Técnicas Electroquímicas , Nanopartículas del Metal , Nanotubos de Carbono , Platino (Metal) , Nanotubos de Carbono/química , Cafeína/análisis , Cafeína/química , Platino (Metal)/química , Nanopartículas del Metal/química , Técnicas Electroquímicas/métodos , Electrodos , Límite de Detección , Reproducibilidad de los Resultados , Oxidación-ReducciónRESUMEN
Sodium-ion batteries (SIBs) operating in aqueous electrolyte are an emerging technology that promises to be safer, cheaper, more sustainable and more efficient than their lithium-based counterparts. One of the great challenges associated with this technology is the development of advanced materials with high specific capacity to be used as electrodes. Herein, we describe an ingenious strategy to prepare unprecedented tri-component nanoarchitected thin films with superior performance when applied as anodes in aqueous SIBs. Taking advantage of the broadness and versatility of the liquid-liquid interfacial route, three transparent nanocomposite films comprising graphene, molybdenum sulphide and copper oxide nanoparticles have been prepared. The samples were characterized using several techniques, and the results demonstrated that depending on the specific experimental strategy, different nanoarchitectures are achieved, resulting in different and improved properties. An astonishing capacity of 1377 mA h g-1 at 0.1 A g-1 and a degree of recovery of 100% were observed for the film in which the interactions among the components were optimized. This is among the highest capacity values reported in the literature and demonstrates the potential of these tri-component materials to be used as anodes in aqueous sodium-ion batteries.
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The appearance of new viruses and diseases has made the development of rapid and reliable diagnostic tests crucial. In light of it, we proposed a new method for assembling an electrochemical immunosensor, based on a one-step approach for selective layer formation. For this purpose, a mixture containing the immobilizing agent (polyxydroxybutyrate, PHB) and the recognition element (antibodies against SARS-CoV-2 nucleocapsid protein) was prepared and used to modify a screen-printed carbon electrode with electrodeposited graphene oxide, for the detection of SARS-CoV-2 nucleocapsid protein (N-protein). Under optimum conditions, N-protein was successfully detected in three different matrixes - saliva, serum, and nasal swab, with the lowest detectable values of 50 pg mL-1, 1.0 ng mL-1, and 50 pg mL-1, respectively. Selectivity was assessed against SARS-CoV-2 receptor-binding domain protein (RBD) and antibodies against yellow fever (YF), and no significant response was observed in presence of interferents, reinforcing the suitability of the proposed one-step approach for selective layer formation. The proposed biosensor was stable for up to 14 days, and the mixture was suitable for immunosensor preparation even after 60 days of preparation. The proposed assembly strategy reduces the cost, analysis time, and waste generation. This reduction is achieved through miniaturization, which results in the decreased use of reagents and sample volumes. Additionally, this approach enables healthcare diagnostics to be conducted in developing regions with limited resources. Therefore, the proposed one-step approach for selective layer formation is a suitable, simpler, and a reliable alternative for electrochemical immunosensing.
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Técnicas Biosensibles , COVID-19 , Humanos , COVID-19/diagnóstico , Inmunoensayo , SARS-CoV-2 , Anticuerpos , Proteínas de la NucleocápsideRESUMEN
This study presents a novel approach for the simultaneous detection of ascorbic acid (AA) and dopamine (DA) using an affordable and user-friendly microfluidic device. Microfluidic devices, when combined with electrochemical detectors like screen-printed electrodes (SPEs), offer numerous advantages such as portability, high sample throughput, and low reagent consumption. In this study, a 3D-printed microfluidic device called a µTED was developed, utilizing textile threads as microfluidic channels and an unmodified SPE as the amperometric detector. The method employed multiple pulse amperometry (MPA) with carefully selected potential values (+0.65 V and -0.10 V). The reduction current signals generated by dopamine o-quinone were used to calculate a correction factor for the oxidation signals of ascorbic acid, enabling simultaneous quantification. The developed microfluidic device ensured a stable flow rate of the carrier solution at 1.19 µL s-1, minimizing the consumption of samples and reagents (injection volume of 2.0 µL). Under the optimized experimental conditions, a linear range from 50 to 900 µmol L-1 was achieved for both DA and AA. The obtained sensitivities were 2.24 µA L mmol-1 for AA and 5.09 µA L mmol-1 for DA, with corresponding limits of detection (LOD) of 2.60 µmol L-1 and 1.54 µmol L-1, respectively. To confirm the effectiveness of the proposed method, it was successfully applied to analyze AA and DA in a commercial blood serum sample spiked at three different concentration levels, with a medium recovery rate of 70%. Furthermore, the MPA technique demonstrated its simplicity by enabling the simultaneous determination of AA and DA without the need for prior separation steps or the use of chemically modified electrodes.
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Dopamina , Microfluídica , Ácido Ascórbico , Dispositivos Laboratorio en un ChipRESUMEN
Neglected tropical diseases are those caused by infectious agents or parasites and are considered endemic in low-income populations. These diseases also have unacceptable indicators and low investment in research, drug production, and control. Tropical diseases such as leishmaniasis are some of the main causes of morbidity and mortality around the globe. Electrochemical immunosensors are promising tools for diagnostics against these diseases. One such benefit is the possibility of assisting diagnosis in isolated regions, where laboratory infrastructure is lacking. In this work, different peptides were investigated to detect antibodies against Leishmania in human and canine serum samples. The peptides evaluated (395-KKG and 395-G) have the same recognition site but differ on their solid-binding domains, which ensure affinity to spontaneously bind to either graphene oxide (GO) or graphene quantum dots (GQD). Cyclic voltammetry and differential pulse voltammetry were employed to investigate the electrochemical behavior of each assembly step and the role of each solid-binding domain coupled to its anchoring material. The graphene affinity peptide (395-G) showed better reproducibility and selectivity when coupled to GQD. Under the optimized set of experimental conditions, negative and positive human serum samples responses were distinguished based on a cut-off value of 82.5% at a 95% confidence level. The immunosensor showed selective behavior to antibodies against Mycobacterium leprae and Mycobacterium tuberculosis, which are similar antibodies and potentially sources of false positive tests. Therefore, the use of the graphene affinity peptide as a recognition site achieved outstanding performance for the detection of Leishmania antibodies.
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Técnicas Biosensibles , Grafito , Leishmaniasis , Animales , Perros , Humanos , Carbono/química , Grafito/química , Reproducibilidad de los Resultados , Inmunoensayo , Péptidos , Anticuerpos , Leishmaniasis/diagnósticoRESUMEN
3D-printing has shown an outstanding performance for the production of versatile electrochemical devices. However, there is a lack of studies in the field of 3D-printed miniaturized settings for multiplex biosensing. In this work, we propose a fully 3D-printed micro-volume cell containing six working electrodes (WEs) that operates with 250 µL of sample. A polylactic acid/carbon black conductive filament (PLA/CB) was used to print the WEs and subsequently modified with graphene oxide (GO), to support protein binding. Cyclic voltammetry was employed to investigate the electrochemical behaviour of the novel multi-electrode cell. In the presence of K3[Fe(CN)6], PLA/CB/GO showed adequate peak resolution for subsequent label-free immunosensing. The innovative 3D-printed cell was applied for multiplex voltammetric detection of three COVID-19 biomarkers as a proof-of-concept. The multiple sensors showed a wide linear range with detection limits of 5, 1 and 1 pg mL-1 for N-protein, SRBD-protein, and anti-SRBD, respectively. The sensor performance enabled the selective sequential detection of N protein, SRBD protein, and anti-SRBD at biological levels in saliva and serum. In summary, the miniaturized six-electrode cell presents an alternative for the low-cost and fast production of customizable devices for multi-target sensing with promising application in the development of point-of-care sensors.
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COVID-19 , Humanos , COVID-19/diagnóstico , Electrodos , Microelectrodos , Poliésteres , Impresión Tridimensional , BiomarcadoresRESUMEN
To help meet the global demand for reliable and inexpensive COVID-19 testing and environmental analysis of SARS-CoV-2, the present work reports the development and application of a highly efficient disposable electrochemical immunosensor for the detection of SARS-CoV-2 in clinical and environmental matrices. The sensor developed is composed of a screen-printed electrode (SPE) array which was constructed using conductive carbon ink printed on polyethylene terephthalate (PET) substrate made from disposable soft drink bottles. The recognition site (Spike S1 Antibody (anti-SP Ab)) was covalently immobilized on the working electrode surface, which was effectively modified with carbon black (CB) and gold nanoparticles (AuNPs). The immunosensing material was subjected to a multi-technique characterization analysis using X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) with elemental analysis via energy dispersive spectroscopy (EDS). The electrochemical characterization of the electrode surface and analytical measurements were performed using cyclic voltammetry (CV) and square-wave voltammetry (SWV). The immunosensor was easily applied for the conduct of rapid diagnoses or accurate quantitative environmental analyses by setting the incubation period to 10 min or 120 min. Under optimized conditions, the biosensor presented limits of detection (LODs) of 101 fg mL-1 and 46.2 fg mL-1 for 10 min and 120 min incubation periods, respectively; in addition, the sensor was successfully applied for SARS-CoV-2 detection and quantification in clinical and environmental samples. Considering the costs of all the raw materials required for manufacturing 200 units of the AuNP-CB/PET-SPE immunosensor, the production cost per unit is 0.29 USD.
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Electrochemical immunosensors are excellent alternatives to prepare portable platforms used for rapid and inexpensive diagnostic of infectious diseases such as the recently emerged COVID-19. Incorporating synthetic peptides as selective recognition layers combined with nanomaterials such as gold nanoparticles (AuNPs) can significantly enhance the analytical performance of immunosensors. In the present study, an electrochemical immunosensor based on solid-binding peptide was built and evaluated towards SARS-CoV-2 Anti-S antibodies detection. The peptide used as recognition site has two important portions: one based on the viral receptor binding domain (RBD), capable of recognizing antibodies of the spike protein (Anti-S), and another suitable for interacting with gold nanoparticles. Gold-binding peptide (Pept/AuNP) dispersion was used directly to modify a screen-printed carbon electrode (SPE). The voltammetric behavior of the [Fe(CN)6]3-/4- probe after every construction and detection step was recorded using cyclic voltammetry by assessing the stability of the Pept/AuNP as a recognition layer onto the electrode surface. Differential pulse voltammetry was used as a detection technique, and a linear working range from 75 ng mL-1 to 15 µg mL-1 was established, with 1.059 µA dec-1 of sensitivity and R2 = 0.984. The response selectivity against SARS-CoV-2 Anti-S antibodies was investigated in presence of concomitant species. The immunosensor was used to detect SARS-CoV-2 Anti-spike protein (Anti-S) antibodies in human serum samples, successfully differentiating between negative and positive responses of samples at a 95% confidence level. Therefore, the gold-binding peptide is a promising tool to be applied as a selective layer for antibody detection.
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Técnicas Biosensibles , COVID-19 , Nanopartículas del Metal , Humanos , Oro/química , SARS-CoV-2 , Técnicas Biosensibles/métodos , Nanopartículas del Metal/química , Inmunoensayo/métodos , Anticuerpos Antivirales , Péptidos , Técnicas Electroquímicas/métodosRESUMEN
The use of biological components in the development of new methods of analysis and point-of-care (POC) devices is an ever-expanding theme in analytical chemistry research, due to the immense potential for early diagnosis of diseases and monitoring of biomarkers. In the present work, the evaluation of an electrochemical microfluidic device based on the immobilization of horseradish peroxidase (HRP) enzyme into chemically treated cotton threads is described. This bioreactor was used as a channel for the build of the microfluidic device, which has allowed to use of a non-modified screen-printed electrode (SPE) as an amperometric detector. Cotton threads were treated using citric acid, and the immobilization of HRP has been performed by EDC/NHS crosslinking, connecting amine groups of the enzymes to carboxylic acids in the cellulosic structure. For the analytical evaluation, an amperometric assay for hydrogen peroxide detection was performed after the injection of H2O2 and hydroquinone (HQN) concomitantly. The enzymatic reaction consumes H2O2 leading to the formation of O-quinone, which is readily reducible at non-modified SPE. Several experimental parameters related to enzyme immobilization have been investigated and under the best set of conditions, a good analytical performance was obtained. In addition, the threads were freezer-stored and, after 12 weeks, 84% of hydrogen peroxide sensitivity was maintained, which is very reasonable for enzyme-based systems and still offers good analytical precision. Therefore, a simple and inexpensive microfluidic system was reported by crosslinking carboxylic groups to amine-containing macromolecules, suggesting a new platform for many other protein-based assays.
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Técnicas Biosensibles , Peróxido de Hidrógeno , Peroxidasa de Rábano Silvestre/química , Peróxido de Hidrógeno/química , Microfluídica , Técnicas Biosensibles/métodos , Enzimas Inmovilizadas/química , Pruebas de Enzimas , AminasRESUMEN
Waste management is a key feature to ensure sustainable consumption and production patterns, and to combat the impacts of climate change. In this scenario, the production of biochar from different biomasses results in environmental and economic advantages. In this study, biochar was produced from sugarcane bagasse pyrolysis, to immobilize biomolecules, in order to assemble an electrochemical immunosensor to detect antibodies against SARS-CoV-2. For this, screen-printed carbon electrodes (SPCE) were modified with a dispersion of biochar and used to immobilize the receptor-binding-domain (RBD) against virus S-protein, through EDC/NHS crosslinking reaction. Under the best set of experimental conditions, negative and positive serum samples responses distinguished based on a cutoff value of 82.3 %, at a 95 % confidence level. The immunosensor showed selective behavior to antibodies against yellow fever and its performance was stable up to 7 days of storage. Therefore, biochar yielded from sugarcane bagasse is an ecofriendly material that can be used as a platform to immobilize biomolecules for construction of electrochemical biosensors.
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Técnicas Biosensibles , COVID-19 , Saccharum , Técnicas Electroquímicas/métodos , SARS-CoV-2 , Celulosa , Inmunoensayo/métodos , Electrodos , AnticuerposRESUMEN
The development of immunosensors to detect antibodies or antigens has stood out in the face of traditional methods for diagnosing emerging diseases such as the one caused by the SARS-CoV-2 virus. The present study reports the construction of a simplified electrochemical immunosensor using a graphene-binding peptide applied as a recognition site to detect SARS-CoV-2 antibodies. A screen-printed electrode was used for sensor preparation by adding a solution of peptide and reduced graphene oxide (rGO). The peptide-rGO suspension was characterized by scanning electron microscopy (SEM), Raman spectroscopy, and Fourier transform infrared spectroscopy (FT-IR). The electrochemical characterization (electrochemical impedance spectroscopy-EIS, cyclic voltammetry-CV and differential pulse voltammetry-DPV) was performed on the modified electrode. The immunosensor response is based on the decrease in the faradaic signal of an electrochemical probe resulting from immunocomplex formation. Using the best set of experimental conditions, the analytic curve obtained showed a good linear regression (r2 = 0.913) and a limit of detection (LOD) of 0.77 µg mL-1 for antibody detection. The CV and EIS results proved the efficiency of device assembly. The high selectivity of the platform, which can be attributed to the peptide, was demonstrated by the decrease in the current percentage for samples with antibody against the SARS-CoV-2 S protein and the increase in the other antibodies tested. Additionally, the DPV measurements showed a clearly distinguishable response in assays against human serum samples, with sera with a response above 95% being considered negative, whereas responses below this value were considered positive. The diagnostic platform developed with specific peptides is promising and has the potential for application in the diagnosis of other infections that lead to high antibody titers.
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Técnicas Biosensibles , COVID-19 , Grafito , Humanos , Grafito/química , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , SARS-CoV-2 , Espectroscopía Infrarroja por Transformada de Fourier , Inmunoensayo , COVID-19/diagnóstico , Electrodos , Límite de Detección , PéptidosRESUMEN
In the present work, we report an innovative approach for immunosensors construction. The experimental strategy is based on the anchoring of biological material at screen-printed carbon electrode (SPE) modified with electrodeposited Graphene Quantum Dots (GQD) and polyhydroxybutyric acid (PHB). It was used as functional substract basis for the recognition site receptor-binding domain (RBD) from coronavirus spike protein (SARS-CoV-2), for the detection of Anti-S antibodies (AbS). SEM images and EDS spectra suggest an interaction of the protein with GQD-PHB sites at the electrode surface. Differential pulse voltametric (DPV) measurements were performed before and after incubation, in presence of the target, shown a decrease in voltametric signal of an electrochemical probe ([Fe(CN)6]3/4-). Using the optimal experimental conditions, analytical curves were performed in PBS and human serum spiked with AbS showing a slight matrix effect and a relationship between voltametric signal and AbS concentration in the range of 100 ng mL-1 and 10 µg mL-1. The selectivity of the proposed sensor was tested against yellow fever antibodies (YF) and the selective layer on the electrode surface did not interact with these unspecific antibodies. Eight samples of blood serum were analyzed and 87.5% of these total investigated provided adequate results. In addition, the present approach showed better results against traditional EDC/NHS reaction with enhancements in time and the possibility to develop an immunosensor in a single drop, since the proteins can be anchored prior to the electrode modification step.
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Técnicas Biosensibles , COVID-19 , Grafito , Puntos Cuánticos , Humanos , Grafito/química , Puntos Cuánticos/química , SARS-CoV-2 , Técnicas Electroquímicas/métodos , Glicoproteína de la Espiga del Coronavirus , Límite de Detección , Inmunoensayo , Electrodos , Carbono/química , AnticuerposRESUMEN
A low-cost and disposable graphene polylactic (G-PLA) 3D-printed electrode modified with gold particles (AuPs) was explored to detect the cDNA of SARS-CoV-2 and creatinine, a potential biomarker for COVID-19. For that, a simple, non-enzymatic electrochemical sensor, based on a Au-modified G-PLA platform was applied. The AuPs deposited on the electrode were involved in a complexation reaction with creatinine, resulting in a decrease in the analytical response, and thus providing a fast and simple electroanalytical device. Physicochemical characterizations were performed by SEM, EIS, FTIR, and cyclic voltammetry. Square wave voltammetry was employed for the creatinine detection, and the sensor presented a linear response with a detection limit of 0.016 mmol L-1. Finally, a biosensor for the detection of SARS-CoV-2 was developed based on the immobilization of a capture sequence of the viral cDNA upon the Au-modified 3D-printed electrode. The concentration, immobilization time, and hybridization time were evaluated in presence of the DNA target, resulting in a biosensor with rapid and low-cost analysis, capable of sensing the cDNA of the virus with a good limit of detection (0.30 µmol L-1), and high sensitivity (0.583 µA µmol-1 L). Reproducible results were obtained (RSD = 1.14%, n = 3), attesting to the potentiality of 3D-printed platforms for the production of biosensors.
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Técnicas Biosensibles , COVID-19 , Grafito , COVID-19/diagnóstico , Creatinina , ADN Complementario , Técnicas Electroquímicas/métodos , Electrodos , Grafito/química , Humanos , Poliésteres , Impresión Tridimensional , SARS-CoV-2RESUMEN
A novel electrochemical sensor based on activated biochar (AB4) and reduced graphene oxide (rGO) was developed and tested for detection of paraquat (PQ) in food samples. Precursor biochar was obtained by the pyrolysis of water hyacinth biomass at 400, 500, and 600 °C, followed by a chemical activation step using HNO3 to increase the amount of oxygenated and nitrogenated groups. The modified electrodes (rGO-AB4) were tested in different experimental conditions, and exhibited good response under the optimized conditions, showing linearity from 0.74 to 9.82 µmol L-1 and a limit of detection and limit of quantification of 0.02 µmolL-1 and 0.07 µmol L-1, respectively. Interfering species such as glyphosate caused insignificant changes in the peak current of paraquat, and the selectivity of the method was tested using blank and spiked samples of coconut water, wastewater, honey, lettuce and lemon. Recovery ranged from 87.70±2.07% to 103.80±3.94%.
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Grafito , Nanocompuestos , Técnicas Electroquímicas , Electrodos , Límite de Detección , ParaquatRESUMEN
Human immunodeficiency virus (HIV) is still considered a pandemic, and the detection of p24-HIV protein has an important role in the early diagnosis of HIV in adults and newborns. The accessibility of these trials depends on the price and execution difficulty of the method, which can be reduced using electrochemical methods by using enzymeless approaches, disposable and accurate devices. In this work, graphene quantum dots were acquired by a simple synthesis and employed as an electrochemical signal amplifier and support for the aptamer immobilization through a feasible and stable modification of disposable screen-printed electrodes. The device has been easily assembled and used to detect p24-HIV protein without the interference of similar proteins or sample matrix. Using the best set of experimental conditions, a linear correlation between analytical signal and log of p24-HIV concentration from 0.93 ng mL-1 to 93 µg mL-1 and a limit of detection of 51.7 pg mL-1 were observed. The developed device was applied to p24 determination in spiked human serum and provided distinct levels of signal for positive and negative samples, successfully identifying real samples with the target protein. This sensor is a step towards the development of point-of-care devices and the popularization of electrochemical methods for trials and diagnostics of relevant diseases.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Grafito , Infecciones por VIH , Puntos Cuánticos , Adulto , Técnicas Electroquímicas , Electrodos , Infecciones por VIH/diagnóstico , Proteínas del Virus de la Inmunodeficiencia Humana , Humanos , Recién Nacido , Límite de DetecciónRESUMEN
Simple, low-cost, and sensitive new platforms for electrochemical immunosensors for virus detection have been attracted attention due to the recent pandemic caused by a new type of coronavirus (SARS-CoV-2). In the present work, we report for the first time the construction of an immunosensor using a commercial 3D conductive filament of carbon black and polylactic acid (PLA) to detect Hantavirus Araucaria nucleoprotein (Np) as a proof-of-concept. The recognition biomolecule was anchored directly at the filament surface by using N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-Hydroxysuccinimide (EDC/NHS). Conductive and non-conductive composites of PLA were characterized using thermal gravimetric analysis (TGA), revealing around 30% w/w of carbon in the filament. Morphological features of composites were obtained from SEM and TEM measurements. FTIR measurement revealed that crosslinking agents were covalently bonded at the filament surface. Electrochemical techniques such as cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used for the evaluation of each step involved in the construction of the proposed immunosensor. The results showed the potentiality of the device for the quantitative detection of Hantavirus Araucaria nucleoprotein (Np) from 30 µg mL-1 to 240 µg mL-1 with a limit of detection of 22 µg mL-1. Also, the proposed immunosensor was applied with success for virus detection in 100x diluted human serum samples. Therefore, the PLA conductive filament with carbon black is a simple and excellent platform for immunosensing, which offers naturally carboxylic groups able to anchor covalently biomolecules.
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Anticuerpos Antivirales/inmunología , Inmunoensayo/métodos , Proteínas de la Nucleocápside/inmunología , Impresión Tridimensional , Anticuerpos Inmovilizados/química , Anticuerpos Inmovilizados/inmunología , COVID-19/diagnóstico , COVID-19/virología , Espectroscopía Dieléctrica , Electrodos , Orthohantavirus/aislamiento & purificación , Orthohantavirus/metabolismo , Infecciones por Hantavirus/diagnóstico , Infecciones por Hantavirus/virología , Humanos , Inmunoensayo/instrumentación , Límite de Detección , Proteínas de la Nucleocápside/sangre , SARS-CoV-2/aislamiento & purificación , Hollín/químicaRESUMEN
The detection of phenolic compounds is relevant not only for their possible benefits to human health but also for their role as chemical pollutants, including as endocrine disruptors. The required monitoring of such compounds on-site or in field analysis can be performed with electrochemical biosensors made with polyphenol oxidases (PPO). In this review, we describe biosensors containing the oxidases tyrosinase and laccase, in addition to crude extracts and tissues from plants as enzyme sources. From the survey in the literature, we found that significant advances to obtain sensitive, robust biosensors arise from the synergy reached with a diversity of nanomaterials employed in the matrix. These nanomaterials are mostly metallic nanoparticles and carbon nanostructures, which offer a suitable environment to preserve the activity of the enzymes and enhance electron transport. Besides presenting a summary of contributions to electrochemical biosensors containing PPOs in the last five years, we discuss the trends and challenges to take these biosensors to the market, especially for biomedical applications.
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Técnicas Biosensibles , Monofenol Monooxigenasa , Nanoestructuras , Carbono , Técnicas Electroquímicas , LacasaRESUMEN
For the first time, a nanocomposite based on biochar and reduced graphene oxide (rGO) was employed to construct a modified carbon paste electrode and applied for the determination of carbendazim (CBZ). Biochar was obtained by through pyrolysis of Eichhornia crassipes biomass, also known how "Aguapé" at 400 °C. The modified electrode with our nanocomposite proposal shows to be able to preconcentrate CBZ and presented the highest analytical response in comparison to the unmodified electrode and by the electrodes prepared with the proposed materials separately. Using differential pulse voltammetry (DPV) under optimized conditions, the sensor showed a linear dynamic response (LDR) from 30 to 900 nmol L-1, a limit of detection (LOD) of 2.3 nmol L-1 and limit of quantification (LOQ) of 7.7 nmol L-1. No significant influence of inorganic ions or organic compounds on sensor response was verified, considering the recovery evaluation data. The proposed sensor was successfully applied for the determination of CBZ in spiked whole orange juice, lettuce leaves, drinking water, and wastewater samples. Good recovery values were found using the ex-situ methodology, showing excellent analytical performance of the electrochemical sensor based on biochar and rGO nanocomposite.