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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: [email protected]

Received date: May 21, 2013; Accepted date: June 12, 2013; Published date: 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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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).

pharmaceutica-analytica-acta-xanthine-oxidase

Figure 1: Generation of superoxide. (.O2 -) and hydrogen peroxide (H2O2) from hypoxanthine and xanthine oxidase.

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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