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Flow Injection Analysis of Hydrogen Peroxide Using Immobilized Horseradish Peroxidase and Its Fluorogenic Substrate 3-(p-Hydroxyphenyl)propionic Acid

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Abstract

A highly sensitive, rapid and precise method for the determination of hydrogen peroxide is described which employs flow injection analysis with fluorescence detection. Hydrogen peroxide solution is injected into a water stream, mixed with a solution of 3-(p-hydroxyphenyl)propionic acid, a fluorogenic substrate for horseradish peroxidase. The mixture is then passed through an immobilized horseradish peroxidase column prepared by coupling of the periodate-treated enzyme with aminopropyl glass beads. The fluorescent product in the stream is monitored with a conventional fluorimeter. A sampling rate of 120/h can be attained. The detection limit is 0.5 pmol in a 20-μl injection volume. The immobilized enzyme column can be used for a year or longer without loss of the activity.

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Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Kyushu University, Maidashi, Fukuoka, 812, Japan

    Yohji Hayashi, Kiyoshi Zaitsu & Yosuke Ohkura

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  1. Yohji Hayashi
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  2. Kiyoshi Zaitsu
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  3. Yosuke Ohkura
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Hayashi, Y., Zaitsu, K. & Ohkura, Y. Flow Injection Analysis of Hydrogen Peroxide Using Immobilized Horseradish Peroxidase and Its Fluorogenic Substrate 3-(p-Hydroxyphenyl)propionic Acid. ANAL. SCI. 1, 65–68 (1985). https://doi.org/10.2116/analsci.1.65

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  • DOI: https://doi.org/10.2116/analsci.1.65

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