Flow Injection Analysis of Glycerol Based on Oxygen Consumption Using Glycerol Dehydrogenase and NADH Oxidase.

  • UKEDA Hiroyuki
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • FUJITA Yoshihiro
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • SAWAMURA Masayoshi
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • KUSUNOSE Hirozo
    Department of Bioresources Science, Faculty of Agriculture, Kochi University

Abstract

A flow injection analysis system for the determination of glycerol has been developed which incorporates a Clark-type oxygen electrode and an enzyme reactor packed with Sepharose 4B on which glycerol dehydrogenase and NADH oxidase are co-immobilized. The response of the system depended on the concentration of flavine adenine dinucleotide (FAD) added into the carrier solution [0.1M glycylglycine-KOH buffer (pH 9.2) containing 30mM ammonium sulfate]. A linear relationship between the glycerol concentration and the response current in the presence of 60μM FAD was observed over the 0.1-1.5mM range. The sampling frequency was about 25 samples/h at a flow rate of 0.5ml/min, and the relative standard deviation was 0.65% at 2.5mM glycerol level (n=10). Application of this system to commercial white and red wines gave satisfactory results compared with the F-kit method. No sample pretreatment was necessary other than dilution.

Journal

  • Analytical Sciences

    Analytical Sciences 10 (3), 445-448, 1994

    The Japan Society for Analytical Chemistry

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