Simultaneous Determination of D-Glucose and 3-Hydroxybutyrate by Chemiluminescence Detection with Immobilized Enzymes in a Flow Injection System

Abstract

A chemiluminometric flow-through sensor for the simultaneous determination of glucose (Glu) and 3-hydroxybutyrate (HB) in a single sample has been developed. Coimmobilized 3-hydroxybutyrate dehydrogenase/NADH oxidase/peroxidase, a support material, and coimmobilized glucose dehydrogenase/NADH oxidase/peroxidase were packed sequentially in a transparent PTFE tube. The tube was then placed in front of a photomultiplier tube as a flow cell. A two-peak recording was obtained by one injection of the sample solution. The peak heights of the first and second peaks were dependent on the concentrations of HB and Glu, respectively. The calibration graphs for HB and Glu were linear at 0.05 - 10 and 0.1 - 30 μM, respectively. The maximum sample throughput was 30 h^-1. The sensor was stable for two weeks.