RESUMO
An enzyme-amperometric detector cell is described for flow analysis of glutamate in dialysate emerging from an implanted microdialysis probe. Its small size allows it to be placed within a few centimetres of the animal preparation, reducing the delay for data acquisition to around 2 min. The selectivity is provided by glutamate oxidase, immobilised with glutaraldehyde on surfaces adjacent to the 3-electrode system. A film of 1,2-diaminobenzene, electropolymerized on the platinum working electrode, eliminates interference from ascorbic acid and other endogenous electroactive compounds. The high sensitivity (< 0.5 mumol/l) and fast response time of the cell (90% of maximum response in 30 s) make it particularly suitable for investigating conditions that produce rapid changes in brain extracellular glutamate. This is illustrated by monitoring changes in extracellular glutamate subsequent to cardiac arrest, and K(+)-induced local depolarization.
Assuntos
Química Encefálica/fisiologia , Espaço Extracelular/metabolismo , Ácido Glutâmico/metabolismo , Aminoácido Oxirredutases , Animais , Técnicas Biossensoriais , Química Encefálica/efeitos dos fármacos , Enzimas Imobilizadas , Espaço Extracelular/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Masculino , Microdiálise , Oxirredução , Consumo de Oxigênio/fisiologia , Cloreto de Potássio/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
Potentiometric and amperometric measurements were made with microbial fuel cells containing E. coli or yeast as the anodic reducing agent and glucose as the oxidizable substrate. The catalytic effects of thionine and resorufin on the anode reaction were investigated. Results on the potentiometry, polarization, and coulombic output of the cells support a mediator-coupled mechanism for the transfer of electrons from the organism to the electrode in preference to a mechanism of "direct" electrochemical oxidation of glucose or its degradation products. Experiments with (14)C-labeled glucose show that when a microbial fuel cell produces a current under load, exogenous glucose is metabolized to produce (14)CO(2). The Coulombic yields of the cells indicate a high degree of energy conversion in these systems.