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

摘要

A combination technique of some kinds of enzymatic reactions for flow injection analysis with an immobilized enzyme-reactorcolumn 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 determinea 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-sensitivityregion of the detector, and from 0.5 mg dm-3 to 30 g 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.