COMPARISON OF 4 INDIRECT METHODS FOR FLUID SUPEROXIDE-DISMUTASE ACTIVITIES

被引:10
作者
DISILVESTRO, RA
DAVID, C
DAVID, EA
机构
[1] Department of Foods and Nutrition, Purdue University, West Lafayette
关键词
SUPEROXIDE DISMUTASE; PLASMA; SYNOVIAL FLUID;
D O I
10.1016/0891-5849(90)90129-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Relatively small sample dilutions could render fluid extracellular (EC) superoxide dismutase (SOD) activity assays more subject to interfering compounds than tissue SOD assays. Highly variable relative SOD activities were obtained when comparing four indirect assays for several fluid samples (human plasma, human synovial fluid, and plasma from healthy or inflamed rats). Analysis of rat plasma fractionated with Sephadex G-150 showed that each assay (three xanthine oxidase based assays plus a modified pyrogallol assay) detected apparent SOD activity almost entirely at the same molecular weight as rat lung EC SOD. However, unfractionated fluid samples caused interferences with the xanthine oxidase based SOD assays, though not with the pyrogallol method. Examples of interference were stimulation of xanthine oxidase activity, color formation without xanthine oxidase, color formation despite excess Cu-Zn SOD addition, and absorbance changes with cyanide inhibition of EC SOD that were above or below blank values. In summary, relative fluid SOD values depended on the assay used, and a modified pyrogallol assay was not subject to several interferences found for three xanthine oxidase based assays of fluid SOD activity.
引用
收藏
页码:507 / 510
页数:4
相关论文
共 15 条
  • [1] Hassan, Biosynthesis and regulation of superoxide dismutases, Free Radic. Biol. Med., 5, pp. 377-385, (1988)
  • [2] Marklund, Human copper-containing superoxide dismutase of high molecular weight, Proc. Natl. Acad. Sci. USA, 79, pp. 7634-7638, (1982)
  • [3] Marklund, Holme, Hellner, Superoxide dismutase in extracellular fluids, Clin. Chim. Acta., 126, pp. 41-51, (1982)
  • [4] Bannister, Calabrese, Assays for superoxide dismutase, Meth. Biochem. Anal., 32, pp. 279-313, (1987)
  • [5] Igari, Kaneda, Horiuchi, Ono, A remarkable increase of superoxide dismutase activity in synovial fluid of patients with rheumatoid arthritis, Clin. Orthoped. Rel. Res., 162, pp. 282-287, (1982)
  • [6] Marklund, Bjelle, Elmqvist, Superoxide dismutase isoenzymes of the synovial fluid in rheumatoid arthritis and in reactive arthritides, Ann. Rheum. Dis., 45, pp. 847-851, (1986)
  • [7] DiSilvestro, Influence of diet and inflammation on serum copper enzymes, Biology of copper complexes, pp. 59-66, (1987)
  • [8] DiSilvestro, Influence of copper intake and inflammation on rat serum superoxide dismutase activity levels, J. Nutr., 118, pp. 474-479, (1988)
  • [9] Sun, Oberley, Li, A simple method for clinical assay of superoxide dismutase, Clin. Chem., 34, pp. 497-500, (1988)
  • [10] Oyanagui, Reevaluation of assay methods and establishment of kit for superoxide dismutase activity, Anal. Biochem., 142, pp. 290-296, (1984)