ISSN : 2153-2435
Pharmaceutica Analytica Acta
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Superoxide Generating System

Hideharu Shintani*
Chuo University, School of Science, 1-13-27, Kasuga Bunkyo 112-0003 Tokyo, Japan
Corresponding Author : Hideharu Shintani
Chuo University, School of Science
1-13-27, Kasuga Bunkyo 112-0003 Tokyo, Japan
Tel: +81425922336
E-mail: shintani@mail.hinocatv.ne.jp
Received May 21, 2013; Accepted June 12, 2013; Published June 15, 2013
Citation: Shintani H (2013) Superoxide Generating System. Pharm Anal Acta S1:002. doi:10.4172/2153-2435.S1-002
Copyright: © 2013 Shintani H. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Abstract

-

Keywords
Csuperoxide anion radical; Hydrogen peroxide
Introduction
There are several chemical ways of generating superoxide anion radical (.O2-); in the biochemical and medical research fields an enzymatic method using xanthine oxidase is widely used (Figure 1).
Protocol
Reagents used are Hypoxanthine solution (1 mM) and xanthine oxidase, Fenton reaction.
Procedure
1. Take hypoxanthine solution (1 mM). For cell culture, use phosphate-buffered saline or foetal calf serum-free medium instead of usual culture medium.
2. Add xanthine oxidase (0.01-100 munits/mL).
3. Incubate for an appropriate time determined experimentally.
4. For cell culture, remove the hypoxanthine solution by aspiration and add the culture medium. The cells are harvested when ready.
Comments
If xanthine oxidase is used H2O2 and .O2- will be produced. If transition metal ions or their redox active complexes are also present hydroxyl radical will be produced (Fenton reaction). To remove H2O2 add catalase to the reaction.
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