Amperometric Determination of Galactose, Lactose and Dihydroxyacetone Using Galactose Oxidase in a Flow Injection System with Immobilized Enzyme Reactors and On-Line Dialysis
作者:
Hans Lundbǎck,
Bo Olsson,
期刊:
Analytical Letters
(Taylor Available online 1985)
卷期:
Volume 18,
issue 7
页码: 871-889
ISSN:0003-2719
年代: 1985
DOI:10.1080/00032718508066184
出版商: Taylor & Francis Group
关键词: Flow Injection Analysis;Amperometric Detection;Flow-Through Dialyzer;Immobilized-Enzyme Reactor;Galactose Oxidase;Peroxidase;Galactose;Lactose;Dihydroxyacetone;Hydrogen Peroxide
数据来源: Taylor
摘要:
A flow injection manifold containing a dialyzer and reactors with immobilized galactose oxidase and peroxidase was used for the determination of galactose in urine, lactose in milk and dihydroxyacetone in a biotechnological reaction medium. The hydrogen peroxide which is formed by the galactose oxidase reaction was detected by amperometric reduction of a mediator. The latter had been produced from hydrogen peroxide in a peroxidase catalyzed reaction. The hydrogen peroxide detection step was studied with several mediators and hexacyanoferrate (II) was selected. An ion exchange HPLC procedure was used to purify the galactose oxidase, in particular from catalase, and the kinetics and the selectivity of a reactor containing the immobilized enzyme was investigated. Columns for removal of certain interferents such as ascorbic acid were used in the determination of galactose in urine. The response to galactose standards was linear from the detection limit of 2 μM to 60 mM. The throughput was 45 samples per hour and the relative standard deviation 0.4%.
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