Fluorometric Determination of Glucose by Flow Injection Analysis Using Immobilized Enzyme-Reactor Columns Prepared by Coupling Glucose Oxidase and Peroxidase onto Chitosan Beads

Abstract

A combination technique of some kinds of enzymatic reactions for flow injection analysis with an immobilized enzyme-reactor column was studied. This system consisted of hand-made reactor columns packed with glucose oxidase and horseradish peroxidase immobilized onto chitosan beads, and a fluorometric detector. It was applied for the determination of glucose in some beverages, Japanese Sake and liquor samples. Under the recommended conditions, it could determine a sample within 10 min. The calibration curve for glucose was linear from 20.0 mg dm^–3 to 0.5 mg dm^–3 in the low-sensitivity region of the detector, and from 0.5 mg dm^–3 to 30 μg dm^–3 in the high-sensitivity region of the detector. The detection limit (S/N=10) was 30 μg dm^–3 as the concentration of glucose, and 54 ng as its absolute amount. There was no interference from major substances present in either beverage or serum samples.