Analytical Sciences

Flow-through Chemiluminescence Sensor Using Immobilized Histamine Oxidase from Arthrobacter crystallopoietes KAIT-B-007 and Peroxidase for Selective Determination of Histamine

Yoshinori SEKIGUCHI,* Aya NISHIKAWA,* Hiroko MAKITA,** Akira YAMAMURA,* Kunio MATSUMOTO,*  and Nobutoshi KIBA***

*Department of Applied Chemistry, Faculty of Engineering, Kanagawa Institute of Technology, Shimo-ogino 1030, Atsugi 243-0292, Japan
**Department of Biochemistry, Faculty of Engineering, Yokohama National University, Tokiwadai 79-1, Hodogaya, Yokohama 240-0067, Japan
***Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Yamanashi University, Takeda 4-3-11, Kofu 400-8511, Japan

A flow sensor with immobilized oxidases is proposed for the determination of histamine in fish meat. Chemiluminometric measurement of histamine was based on the luminol reaction with hydrogen peroxide produced by immobilized histamine oxidase (EC 1.4.3.-.) and peroxidase (EC 1.11.1.7.) within a flow cell. Histamine oxidase was found in cells of Arthrobacter crystallopoietes KAIT-B-007 isolated from soil. The oxidase and peroxidase were coimmobilized covalently on tresylated hydrophilic vinyl polymer beads and packed into transparent PTFE; the tubing was used as the flow cell. One assay for histamine was done at intervals of 2 min without carryover. The calibration curve for histamine was linear from 0.1 uM to 50 uM. The response was reproducible within 1.25% of the relative standard deviation for 115-replicate injections of 50 uM histamine. The sensor system was applied to the determination of histamine in fish meat extracts.

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