The development of a generic hydrogen peroxide sensor with application to the detection of glucose

Lina Genovesi, Henrik Pedersen, George H. Sigel

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

A generic sensor for hydrogen peroxide producing metabolites was developed. The sensor consists of a small bore nylon tube fitting closely over two UV-transmitting fibers in an optrode configuration. In the case of glucose sensing, horseradish peroxidase and glucose oxidase were coimmobilized on the inside surface of the nylon tube. Glucose oxidase catalyzes the production of hydrogen peroxide from glucose and horseradish peroxidase catalyzes the reaction of hydrogen peroxide (H2O2) and p-hydroxyphenylacetic acid (HPA) into 6-6’-dihydroxy(1,1′-biphenyl) 3,3′-diacetic acid (DBDA) which is measured by laser-assisted fluorometry. The amount of fluorescing DBDA is directly proportional to the amount of glucose present. Michaelis-Menten kinetics for soluble and immobilized enzymes were established for the H2O2/HPA/HRP system to determine the range of hydrogen peroxide concentrations for which sensor linear response is observed. Sensors were found to be stable at 4°C in pH 7 phosphate buffer for a period of thirty days with no significant loss in enzyme activity. Glucose levels in buffer at physiological pH were measured and sensor linearity was established. Applicability of the sensor to the measurement of glucose levels in serum was established. Further studies will involve in vitro sensor calibration.

Original languageEnglish (US)
Pages (from-to)22-28
Number of pages7
JournalProceedings of SPIE - The International Society for Optical Engineering
Volume990
DOIs
StatePublished - Jan 19 1989

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

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