936 resultados para POTENTIOMETRIC BIOSENSOR
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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A new electrochemical hybridization biosensor protocol without an external indicator is described. The biosensor format involves the immobilization of inosine-substituted (guanine-free) probe onto the carbon paste transducer, and a direct chronopotentiometric detection of the duplex formation by the appearance of the guanine oxidation peak of the target. Such a use of the intrinsic DNA electrochemical response for monitoring hybridization events offers several advantages (over the common use of external indicators), including the appearance of a new peak, a Aat background, or simplicity. A 4 min short hybridization period allows a detection limit around 120 ng/ml. Performance characteristics of the sensor are described along with future prospects. (C) 1998 Elsevier B.V. B.V. All rights reserved.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Fractional factorial design and factorial with center point design were applied to the development of an amperometric biosensor for the detection of the hepatitis C virus. Biomolecules were immobilized by adsorption on graphite electrodes modified with siloxane-poly(propyleneoxide) hybrid matrix prepared using the sol-gel method. Several parameters were optimized, such as the streptavidin concentration at 0.01 mg mL(-1) and 1.0% bovine serum albumin, the incubation time of the electrodes in the complementary DNA solution for 30 minutes and a 1: 1500 dilution of the avidin-peroxidase conjugate, among others. The application of chemometric studies has been efficient, since the best conditions have been established with a restricted number of experiments, indicating the influence of different factors on the system.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Desenvolveu-se um biossensor para ácido L-ascórbico empregando ascorbato oxidase. A enzima foi extraída do mesocarpo de pepino com solução tampão fosfato 0,05 mol L-1, pH 5,8 contendo NaCl 0,5 mol L-1. Após diálise versus solução tampão fosfato 0,05 mol L-1, pH 5,8 a enzima foi imobilizada em rede de nylon através de ligação covalente com glutaraldeído. A membrana foi acoplada em eletrodo de O2 e a reação monitorada pelo consumo de oxigênio a -600 mV em análise em fluxo (solução tampão fosfato 0,05 mol L-1, pH 5,8 como carregador e vazão 0,5 mL min-1). A curva analítica apresentou-se linear entre 1,2x10-4 a 1,0x10-3 mol L-1. O tempo de vida do biossensor foi de 500 análises. Amostras de medicamentos foram analisadas com a metodologia proposta e os resultados comparados com os obtidos com HPLC.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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The characteristics, performance, and application of an electrode, namely, Pt vertical bar Hg vertical bar Hg-2(FF)(2)vertical bar Graphite, where FF stands for flufenamate ion, are described. This electrode responds to FF with sensitivity of (-58.6 +/- 1.2) mV decade(-1) over the range 1.0 x 10(-6) - 1.0 x 10(-2) mol L-1 at pH 6.0-9.0 and a detection limit of 7.1 x 10(-7) mol L-1. The electrode is easily constructed at a relatively low cost with fast response time (within 10-25 s) and can be used for a period of 5 months without any considerable change in its performance characteristics. The proposed electrode displayed good selectivity for flufenamate in the presence of several substances as well as in the presence of some carboxylate and inorganic anions. The electrode was successfully applied to the determination of flufenamic acid in pharmaceuticals and human serum samples.
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The application of adsorptive stripping potentiometry to the reductive detection of nucleic acids at mercury electrodes is reported. Compared to analogous voltammetric stripping modes, constant current potentiometric stripping analysis (PSA) effectively addresses the hydrogen discharge background problem, and hence greatly improves the characteristics of the superimposed cytosine/adenine (CA) reduction peak. Compared to earlier schemes for trace measurements of nucleic acids at mercury or carbon electrodes that rely on anodic signals arising from the guanine residue, convenient quantitation can now be carried out in connection with the cytosine and adenine residues. Variables influencing the adsorptive PSA response are explored and optimized. With five minute accumulation, the detection limits for tRNA, ssDNA and dsDNA are 30 mu g l(-1), 60 mu g l(-1) and 2 mg l(-1), respectively. Such different values reflect the strong dependence of the PSA CA signal upon the nucleic-acid structure. This allows the quantitation of ssDNA or tRNA in the presence of dsDNA, and offers new possibilities for electrochemical studies of DNA structure and interactions.
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A potentiometric sensor constructed from a mixture of 25% (m/m) spinel-type manganese oxide (lambda-MnO2), 50% (m/m) graphite powder and 25% (m/m) mineral oil is used for the determination of lithium ions in a flow injection analysis system. Experimental parameters, such as pH of the carrier solution, flow rate, injection sample volume, and selectivity for Li+ against other alkali and alkaline-earth ions and the response time of this sensor were investigated. The sensor response to lithium ions was linear in the concentration range 8.6 x 10(-5) - 1.0 x 10(-2) mol L-1 with a slope 78.9 +/- 0.3 mV dec(-1) over a wide pH range 7 - 10 (Tris buffer), without interference of other alkali and alkaline-earth metals. For a flow rate of 5.0 mL min(-1) and a injection sample volume of 408.6 muL, the relative standard deviation for repeated injections of a 5.0 x 10(-4) mol L-1 lithium ions was 0.3%.