Analytical Sciences

Fluorometric Determination of Glucose by Flow InjectionAnalysis Using Immobilized Enzyme-Reactor ColumnsPrepared by Coupling Glucose Oxidase andPeroxidase onto Chitosan Beads

Tetsuyuki TANIAI, Akio SAKURAGAWA, and Tadao OKUTANI

Department of Materials and Applied Chemistry, College of Science and Technology, Nihon University,Kanda-surugadai, Chiyoda, Tokyo 101-8308, Japan

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 andhorseradish peroxidase immobilized onto chitosan beads, and a fluorometric detector. It was applied for the determinationof 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. Thedetection limit (S/N=10) was 30 g dm^-3 as the concentration of glucose, and 54 ng as its absolute amount. There was nointerference from major substances present in either beverage or serum samples.

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