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Updated: Jul 31, 2026
Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases
Published on: February 24, 2018
Extraction and analysis of superoxide free radicals (.O2-) from whole mammalian liver
A R Shoaf1, A U Shaikh, R D Harbison
1Health Protection Section, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, NC 27103.
Abstract:
Extraction of whole lobes of normal rat liver with dimethyl sulphoxide (DMSO) under N2 gives extracts which contain 5-10 mumol/l.O2- (50-100 nmol.O2- per 10 ml extract per 4 g liver; 1.25-2.50 nmol.O2- per millilitre per gram liver). Evidence for .O2- in the extracts is given by: (1) electron spin resonance signals (ESR), (2) differential pulse polarography (DPP), (3) chemiluminescence (CL), and (4) nitroblue tetrazolium reduction (NBT). All tests yield results identical with those obtained with authentic .O2-. Extraction of .O2- is enhanced by tetrabutyl ammonium ion, and is maximal at 1-3 min. These results raise the possibility that substantial amounts of .O2- are normally sequestered in protective membranous sites in vivo.
