Electrogenerated chemiluminescence of luminol for oxidase-based fibre-optic biosensors

The luminol electrochemiluminescence has been exploited for the development of several fibre‐optic biosensors allowing the detection of hydrogen peroxide and of substrates of H2O2‐producing oxidases. Electro‐optical flow injection analysis of glucose, lactate, cholesterol and choline are thus descri...

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Bibliographic Details
Published in:Luminescence (Chichester, England) England), 2001-03, Vol.16 (2), p.159-165
Main Authors: Marquette, Christophe A., Leca, Béatrice D., Blum, Loïc J.
Format: Article
Language:English
Subjects:
Alcohol Oxidoreductases - metabolism
Biosensing Techniques
Collagen - metabolism
electrochemiluminescence
Electrochemistry
Enzymes, Immobilized - chemistry
Fiber Optic Technology
fibreoptic biosensor
flow injection analysis
Hydrogen Peroxide - metabolism
immobilized enzymes
Luminescent Measurements
luminol
Luminol - metabolism
Membranes, Artificial
Substrate Specificity
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Summary:The luminol electrochemiluminescence has been exploited for the development of several fibre‐optic biosensors allowing the detection of hydrogen peroxide and of substrates of H2O2‐producing oxidases. Electro‐optical flow injection analysis of glucose, lactate, cholesterol and choline are thus described. To perform the experiments, a glassy carbon electrode was polarized at a fixed potential. Luminol was then electrochemically oxidized and could react in the presence of hydrogen peroxide to produce light. Several parameters had to be optimized to obtain reliable optical biosensors. An optimum applied potential of +425 mV between the glassy carbon electrode and the platinum pseudo‐reference electrode was determined, allowing the best signal: noise ratio to be obtained. It was also necessary to optimize the experimental conditions for the immobilization of the different oxidases involved (preactivated membranes, chemically activated collagen membranes, photopolymerized matrix). For each biosensor developed, the optimum reaction conditions have been studied: buffer composition, pH, temperature, flow rate and luminol concentration. Under optimal conditions, the detection limits (S/N = 3) were 30 pmol, 60 pmol, 0.6 nmol and 10 pmol for lactate, glucose, cholesterol and choline, respectively. The miniaturization of electrochemiluminescence‐based biosensors has been realized using screen‐printed electrodes instead of a glassy carbon macroelectrode, with choline oxidase as a model H2O2‐generating oxidase. Copyright © 2001 John Wiley & Sons, Ltd.
ISSN:1522-7235
1522-7243
DOI:10.1002/bio.617