Microchimica Acta

, Volume 121, Issue 1–4, pp 237–247 | Cite as

Fibre optic fluorometric enzyme sensors for hydrogen peroxide and lactate, based on horseradish peroxidase and lactate oxidase

  • Florian Schubert
  • Fang Wang
  • Herbert Rinneberg
Original Papers

Abstract

An optical biosensor for the determination of hydrogen peroxide based on immobilized horseradish peroxidase is described. The fluorescence of the dimeric product of the enzyme catalysed oxidation of homovanillic acid is utilized to determine the concentration of H2O2. The membrane-bound enzyme is attached to a bifurcated fibre bundle permitting excitation and detection of the fluorescence by a fluorometer. The response of the sensor is linear from 1 to 130 μM hydrogen peroxide; the coefficient of variation is 3%. The sensor is stable for more than 10 weeks. The operating pH for maximal sensor response is 8.15. This allows the sensor to be used in combination with oxidase reactions producing hydrogen peroxide, as is demonstrated with a co-immobilized lactate oxidase-horseradish peroxidase optode for the determination of L-lactate. The fluorescence intensity of this sensor depends linearly on the concentration of lactate between 3 and 200 μM and a throughput of 10 samples per hour is possible. The precision is in the same range as that of the monoenzyme optode. The lifetime of the bienzyme sensor for lactate is considerably shorter than that of the peroxidase sensor; it is limited by the stability of the immobilized lactate oxidase enzyme. The sensor has been applied to the determination of lactate in control serum.

Key words

optical biosensor enzyme optode hydrogen peroxide horseradish peroxidase lactate determination lactate oxidase 

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Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Florian Schubert
    • 1
  • Fang Wang
    • 1
  • Herbert Rinneberg
    • 1
  1. 1.Group Medical Measuring TechnologyPhysikalisch-Technische BundesanstaltBerlinFederal Republic of Germany
  2. 2.Laboratory for Macromolecule Analysis, Department of Biochemistry, Albert Einstein College of MedicineYeshiva UniversityBronxUSA

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