Optimization of incubation time of protein Tc85 in the construction of biosensor: Is the EIS a good tool?

Electrochemical impedance spectroscopy (EIS) in pH 6.9 phosphate buffer solution was used to investigate each step of the procedure employed to modify a screen-printed electrode (SPE). The SPE was modified with self-assembled monolayers (SAMs) of cystamine (CYS, deposited from 20 mM solution), followed by glutaraldehyde (GA, 0.3 M solution). The Trypanosoma cruzi antigen was immobilized using different deposition times. The influence of incubation time (2-18 h) of protein was also investigated. The topography of modified electrode with this protein was investigated by atomic force microscopy (AFM). Interpretation of impedance data was based on physical and chemical adsorption, and degradation of the layer at high and meddle frequencies, and charge transfer reaction involving mainly the reduction of oxygen at low frequencies. EIS studies on modified electrodes with Tc85 protein immobilized for different incubation times indicated that the optimum incubation time was 6-8 h. It was demonstrated that EIS is a good technique to evaluate the different steps and the integrity of the surface modifications, and to optimize the incubation time of protein in the development of biosensors. (C) 2010 Elsevier B.V. All rights reserved.

Optimization of incubation time of protein Tc85 in the construction of biosensor: Is the EIS a good tool?

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detection of carbamate pesticides in vegetable samples using cholinesterase-based biosensors

Biosensors for determination of carbamates in vegetables based on five different cholinesterases as biorecognition elements and a screen-printed electrode system as an amperometric transducer were developed. Measurements were simply performed by dropping solutions (either sample or substrate) directly on the biosensor. The response of biosensors to selected carbamates (aldicarb, carbaryl, carbofuran, methomyl and propoxur) was characterized. The performance was evaluated on extracts of potatoes and carrots, the results from the AChE biosensor and a standard HPLC procedure were compared. Finally, the biosensor was used for the direct analysis of vegetable juices without any pretreatment steps. In this case, 10 mu g/L levels of added carbofuran and propoxur were reliably identified. The whole procedure takes less than 20 min including 10 min incubation with samples. The concentrations of carbamates determined with biosensor agreed well for carbofuran. Lower response was observed for propoxur.

A disposaable electrochemical sensor for the rapid determination of levodopa

Levodopa (L-dopa), the biological precursor of catecholamines, is the most widely prescribed drug in the treatment of Parkinson's disease. The present work presents a proposal for the application of a gold screen-printed electrode an electrochemical sensor for monitoring L-dopa in stationary solution and a flow system. Using the electrooxidation of L-dopa at +0.63 V in acetate buffer pH 3.0 on a gold screen-printed electrode it is possible to obtain a linear calibration curve from 9.9 x 10(-5) to 1.2 x 10(-3) mol L-1 and a detection limit of 6.8 x 10(-5) mol L-1. Under amperometric conditions (E-app = 0.8 V; flow rate = 14.1 ml, min(-1); pH 3.0), an analytical calibration graph for L-dopa was obtained from 1.0 x 10(-6) mol L-1 6.6 x 10(-4) mol L-1 with a detection limit of 9.9 x 10(-7) mol L-1. The method was successfully applied to the determination of L-dopa in commercial dosage forms without any pre-treatment. (c) 2005 Elsevier B.V. All rights reserved.

Development of a voltammetric sensor for chromium(VI) determination in wastewater sample

Screen-printed carbon electrode (SPCE) modified with poly-L-histidine film can be successfully applied for chromium(VI) determination based on its pre-concentration. Optimum adherence and stability of the POIY-L-histidine film was obtained by direct addition of PH solution 1% (w/v) on the electrode surface, followed by heating at 80 degrees C during 5 min. Linear response range, sensitivity and limit of detection were 0. 1-150 [mu mol L-1, 4. 13 LA mu mol L` and 0.046 mu mol L-1. The repeatability of the proposed sensor, evaluated in terms of relative standard deviation, was measured as 3.2% for 10 experiments in 40 mu mol L-1 using the same electrode and 4.0% using screen-printed electrode as disposable sensor, respectively. The voltammetric sensor was applied to determination of Cr(VI) and indirect determination of Cr(III) in wastewater samples previously treated by a leather dyeing industry and the average recovery for these samples was around 97%. (C) 2006 Elsevier B.V. All rights reserved.

A novel LTCC electrochemical cell construction and characterization: A detection compartment for portable devices

In this work we described for the first time the construction of a 25 μL electrochemical cell from low temperature co-fired ceramic (LTCC) material and carbon screen-printed electrode applicable in portable devices. Firstly, a carbon screen-printed electrode was prepared and characterized by cyclic voltammetry and scanning electron microscopy. Afterwards carbon polymeric film and metal pastes were dropped into the LTCC cell cavities in order to determine the device electrodes, and this arrangement was also electrochemically characterized. The great advantage of this promising device is the simple construction method and its widespread applicability in reusable portable devices. © 2013 The Royal Society of Chemistry.

Eletrodos impressos descartáveis e de carbono vítreo modificados com grafeno e compósito de nanotubos de carbono-quitosana aplicados na determinação de corantes de cabelo

Pós-graduação em Química - IQ

Análise de acetona em ar exalado: desenvolvimento de método eletroanalítico e algoritmo para processamento de sinais

Propõe-se método novo e completo para análise de acetona em ar exalado envolvendo coleta com pré-concentração em água, derivatização química e determinação eletroquímica assistida por novo algoritmo de processamento de sinais. Na literatura recente a acetona expirada vem sendo avaliada como biomarcador para monitoramento não invasivo de quadros clínicos como diabetes e insuficiência cardíaca, daí a importância da proposta. Entre as aminas que reagem com acetona para formar iminas eletroativas, estudadas por polarografia em meados do século passado, a glicina apresentou melhor conjunto de características para a definição do método de determinação por voltametria de onda quadrada sem a necessidade de remoção de oxigênio (25 Hz, amplitude de 20 mV, incremento de 5 mV, eletrodo de gota de mercúrio). O meio reacional, composto de glicina (2 mol·L-1) em meio NaOH (1 mol·L-1), serviu também de eletrólito e o pico de redução da imina em -1,57 V vs. Ag|AgCl constituiu o sinal analítico. Para tratamento dos sinais, foi desenvolvido e avaliado um algoritmo inovador baseado em interpolação de linha base por ajuste de curvas de Bézier e ajuste de gaussiana ao pico. Essa combinação permitiu reconhecimento e quantificação de picos relativamente baixos e largos sobre linha com curvatura acentuada e ruído, situação em que métodos convencionais falham e curvas do tipo spline se mostraram menos apropriadas. A implementação do algoritmo (disponível em http://github.com/batistagl/chemapps) foi realizada utilizando programa open source de álgebra matricial integrado diretamente com software de controle do potenciostato. Para demonstrar a generalidade da extensão dos recursos nativos do equipamento mediante integração com programação externa em linguagem Octave (open source), implementou-se a técnica da cronocoulometria tridimensional, com visualização de resultados já tratados em projeções de malha de perspectiva 3D sob qualquer ângulo. A determinação eletroquímica de acetona em fase aquosa, assistida pelo algoritmo baseado em curvas de Bézier, é rápida e automática, tem limite de detecção de 3,5·10-6 mol·L-1 (0,2 mg·L-1) e faixa linear que atende aos requisitos da análise em ar exalado. O acetaldeído, comumente presente em ar exalado, em especial, após consumo de bebidas alcoólicas, dá origem a pico voltamétrico em -1,40 V, contornando interferência que prejudica vários outros métodos publicados na literatura e abrindo possibilidade de determinação simultânea. Resultados obtidos com amostras reais são concordantes com os obtidos por método espectrofotométrico, em uso rotineiro desde o seu aperfeiçoamento na dissertação de mestrado do autor desta tese. Em relação à dissertação, também se otimizou a geometria do dispositivo de coleta, de modo a concentrar a acetona num volume menor de água gelada e prover maior conforto ao paciente. O método completo apresentado, englobando o dispositivo de amostragem aperfeiçoado e o novo e efetivo algoritmo para tratamento automático de sinais voltamétricos, está pronto para ser aplicado. Evolução para um analisador portátil depende de melhorias no limite de detecção e facilidade de obtenção eletrodos sólidos (impressos) com filme de mercúrio, vez que eletrodos de bismuto ou diamante dopado com boro, entre outros, não apresentaram resposta.

Determinationofgroundwatermercury(II)contentusingadisposable goldmodified screenprintedcarbonelectrode

Mercury (II)measurementswereperformedthankstoanewlydevelopedelectrochemicalmethodusinga disposablegoldmodified screenprintedcarbonelectrode.Themethodhasawidedynamicrange(1–100 mg/L), agoodaccuracyandalimitofdetectionincompliancewithWHOstandards.Theapplicationofthe methodtoseveralgroundwatersamplesmadeitpossibletoidentify,forthe first time,mercurycontent higherthantherecommendedWHOstandardvalueinagoldminingactivityareainthenorthern partofBurkinaFaso.TheaccuracyoftheassaywascheckedbyICP/MS.

Development of an electrochemical biosensor for the detection of miRNA-155 in Breast Cancer

Breast cancer is one of the most prevalent forms of cancer in women. Despite all recent advances in early diagnosis and therapy, mortality data is not decreasing. This is an outcome of the inexistence of validated serum biomarkers allowing an early prognosis, out coming from the limited understanding of the natural history of the disease. In this context, miRNAs have been attracting a special interest throughout the scientific community as promising biomarkers in the early diagnosis of cancer. In breast cancer, several miRNAs and their levels of expression are significantly different between normal tissue and tissue with neoplasia, as well as between different molecular subtypes of breast cancer, also associated with prognosis. Thus, this these presents a meta-analysis that allows identifying a reliable miRNA biomarker for the early detection of breast cancer. In this, miRNA-155 was identified as the best one and an electrochemical biosensor was developed for its detection in serum samples. The biosensor was assembled by following three button-up stages: (1) the complementary miRNA sequence thiol terminated (anti-miRNA-155) was immobilized on a commercial gold screen-printed electrode (Au-SPE), followed by (2) blocking non-specific binding with mercaptosuccinic acid and by (3) miRNA hybridization. The biosensor was able to detect miRNA concentrations lying in the 10-18 mol/L (aM) range, displaying a linear response from 10 aM to 1nM. The device showed a limit of detection of 5.7 aM in human serum samples and good selectivity against other biomolecules in serum, such as cancer antigen CA-15.3 and bovine serum albumin (BSA). Overall, this simple and sensitive strategy is a promising approach for the quantitative and/or simultaneous analysis of multiple miRNA in physiological fluids, aiming at further biomedical research devoted to biomarker monitoring and point-of-care diagnosis.

Aspects of cathodic stripping voltammetry at the hanging mercury drop electrode and in non-mercury disposable sensors

Cathodic stripping voltammetry (CSV) and accumulation at the hanging mercury drop electrode are reviewed briefly. Proposals in a recent IUPAC technical report are considered. Three recent developments in CSV are discussed: the adaptation of CSV methods developed for use with the hanging mercury drop electrode for use with screen-printed carbon electrodes in disposable sensors, the use of reactive accumulation, and the chemometric use of kinetic methods of determination with pulse methods in CSV.

Evaluation of a highly sensitive amperometric biosensor with low cholinesterase charge immobilized on a chemically modified carbon paste electrode for trace determination of carbamates in fruit, vegetable and water samples

A highly sensitive amperometric biosensor for determination of carbamate pesticides directly in water, fruit and vegetable samples has been evaluated, electrochemically characterized and optimized. The biosensor strip was fabricated in screen printed technique on a ceramic support using silver-based paste for reference electrode, and platinum-based paste for working and auxiliary electrodes. The working electrode was modified by a layer of carbon paste mixed with cobalt(II) phthalocyanine and acetylcellulose. Cholinesterase (ChE) enzymes with low enzymatic charge were immobilized on this layer. The operational simplicity of the biosensor consists in that a small drop (similar to 50 mu l) of substrate or sample is deposited on a horizontally positioned biosensor strip representing the microelectrochemical cell. The working potential of the biosensor was 370 mV versus Ag/AgI on a ship reference electrode preventing the interference of electroactive species which are oxidable at more positive potentials. The biosensor was applied to investigate the degradation of two reference ChE inhibitors in freeze dried water under different storage conditions and for direct determination of some N-methylcarbamates (NMCs) in fruit and vegetable samples at ppb concentration levels without any sample pretreatment. A comparison of the obtained results for the total carbamate concentration was done against those obtained using HPLC measurements. (C) 1999 Elsevier B.V. B.V. All rights reserved.

Development of polyaniline microarray electrodes for cadmium analysis

Disposable screen-printed electrodes (SPCE) were modified using a cosmetic product to partially block the electrode surface in order to obtain a microelectrode array. The microarrays formed were electropolymerized with aniline. Scanning electron microscopy was used to evaluate the modified and polymerized electrode surface. Electrochemical characteristics of the constructed sensor for cadmium analysis were evaluated by cyclic and square-wave voltammetry. Optimized stripping procedure in which the preconcentration of cadmium was achieved by depositing at –1.20 V (vs. Ag/AgCl) resulted in a well defined anodic peak at approximately –0.7 V at pH 4.6. The achieved limit of detection was 4 × 10−9 mol dm−3. Spray modified and polymerized microarray electrodes were successfully applied to quantify cadmium in fish sample digests.

Análise voltamétrica da ciprofloxacina – aplicação a medicamentos e a remediação

O presente trabalho tem como objectivo o desenvolvimento de um método analítico, baseado na voltametria de onda quadrada (SWV), para a análise de ciprofloxacina (CIP) em produtos farmacêuticos e em processos de remediação. Para o desenvolvimento do método voltamétrico foram utilizadas duas células voltamétricas: a célula clássica (utilizando um eléctrodo de trabalho de carbono vítreo - GCE) e um eléctrodo de carbono impresso (SPCE). Após a optimização dos parâmetros da SWV, pH (3,04), frequência (400Hz), incremento de potencial (6 mV) e amplitude do impulso de potencial (40 mV), procedeu-se a validação dos métodos, obtendo-se zonas lineares entre a concentração de CIP e a intensidade de corrente de pico de 5,0×10-6 a 6,0×10-5 mol/L (GCE) e de 1,0×10-5 a 4,0×10-5 mol/L (SPCE) e limites de detecção de 9,48×10-6 mol/L (GCE) e 2,13×10-6 mol/L (SPCE). Verificou-se que a sensibilidade, a precisão e a selectividade são superiores para o SPCE, sendo por isso esta a célula mais adequada para proceder à análise da CIP em produtos farmacêuticos. O SPCE foi aplicado com sucesso à análise de CIP num produto farmacêutico. Para o tratamento de soluções aquosas contendo a CIP foram testados dois oxidantes: o permanganato de potássio e o peróxido de hidrogénio. Para o peróxido de hidrogénio os resultados obtidos foram inconclusivos. No caso do permanganato de potássio, os resultados mostram que a degradação da ciprofloxacina depende da concentração do oxidante. Para uma concentração de CIP de 3,00×10-4 mol/L uma degradação rápida foi obtida com o uso de 6,00×10-3 mol/L de permanganato de potássio. Na aplicação do permanganato na remediação de solos verificou-se que no caso de solos húmicos a ciprofloxacina é adsorvida pelo solo, não sendo possível confirmar a ocorrência da reacção de degradação. No caso de solos arenosos verificou-se que a ciprofloxacina foi rapidamente degradada pelo permanganato de potássio.

Nanohybrid materials as transducer surfaces for electrochemical sensing applications

A nanohybrid electrochemical transducer surface was developed using carbon and gold nanomaterials. The strategy relayed on casting multiwalled carbon nanotubes or carbon nanofibers onto a screen-printed carbon electrode surface, followed by in situ generation of gold nanoparticles by electrochemical deposition of ionic gold, in a reproducible manner. These transducers, so fabricated, were characterized using both electrochemical and microscopic techniques. Biofunctionality was evaluated using the streptavidin-biotin interaction system as the biological reaction model. These platforms allow to achieve low detection limits (in the order of pmoles), are reproducible and stable at least for a month after their preparation, being a perfect candidate to be used as transducer of different sensor devices.