©2014 JCBN JCBNournal of Clinical Biochemistry and Nutrition0912-00091880 5 86the Society for Free Radical Rese rch JapanKyot, Japanjcbn13-610.3 64/jcb.13-61Original ArticleScavenging of reactive oxygen species induced

Copyright © 2014 JCBN2014This is an open access article distributed under the terms of theCreat ve Commons Attribution Licens, which permit unre-st icted use, distribution, and reproduction i any medium, p ovided the original work is prop rly cit d.Th scavenging activity of rat plasma against hyper hermia induced reactive oxygen species was tested. The glutathione dependent reduction of a nitroxyl radical, 4hydroxyl2,2,6,6 tetramethylpiperidineNoxyl, which was restricted by adding superoxide dismutase or by deoxygenating the reaction mixture, was applied to an index of superoxide (O2•−) generation. A reaction mixture containing 0.1 mM 4hydroxyl2,2,6,6tetramethylpiperi dineNoxyl and 1 mM glutathione was prepared using 100 mM phosphate buffer containing 0.05 mM diethylenetriaminepenta acetic acid. The reaction mixture was kept in a screwtop vial and incubated in a water bath at 37 or 44°C. The time course of the electron paramagnetic resonance signal of 4hydroxyl2,2,6,6 tetramethylpiperidineNoxyl in the reaction mixture was mea sured by an Xband EPR spectrometer (JEOL, Tokyo, Japan). When the same experiment was performed using rat plasma instead o