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Measurement of Superoxide and Superoxide Dismutase by Electron Spin Resonance and Chemiluminescence Assay

  • Toshikazu Yoshikawa
  • Yuji Naito
  • Toru Tanigawa
  • Shigeru Sugino
  • Motoharu Kondo
Part of the NATO ASI Series book series (NSSA, volume 189)

Abstract

Spin trapping using dimethyl-1-pyrroline-N-oxide (DMPO) s a spin trapper is a useful method for the detection of superoxidel, 2. DMPOO0H which is formed by the reaction of DMPO and superoxide gives a characteristic electron spin resonance (ESR) signal. This combined with its superoxide dismutase (SOD) inhibitability, is making it possible to give a two fold garantee of specificity to this assay. Because of the relatively short life time of DMPO-OOH, to exactly quantify superoxide is difficult. However it is still possible to semi-quantify in cases where the rate of superoxide generation is relatively constant3. These methods may be advantageous when the materials show a strong light absorption or turbidity.

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References

  1. 1).
    Finkelstein E., Rosen G. M., and Rauckman E. J.: Spin trapping of superoxide. Mol. Pharmacol., 16: 676–685, 1979.PubMedGoogle Scholar
  2. 2).
    Finkelstein E., Rosen G. M., and Rauckman E. J.: Spin trapping; kinetics of the reaction of superoxide and hydroxyl radicals with nitrones. J. Am. Chem. Soc. 102: 4994–4999, 1980.CrossRefGoogle Scholar
  3. 3).
    Tanigawa T., Yoshikawa T., Miyagawa H., Ueda S., Takemura T., Tainaka K., Morita Y., Itani K., Yoshida N., Sugino S., and Kondo M.: Determination of superoxide generated by human polymorphonuclear leukocytes by ESR and its clinical application. Jap. J. Inflammation. 8: 443–447, 1988.Google Scholar
  4. 4).
    Sugioka K., Nakano M., Kurashige S., Akuzawa Y., and Goto T.: A chemiluminescence probe with a Cypridana liciferin analog, 2-methyl-6-phenyl-3,7-dihydroimidazo[1,2-a] pyrazin-3-one, specific and sensitive for superoxide production in phagocytizing manrophages. FEBS Lett. 197: 27–30, 1986.CrossRefGoogle Scholar
  5. 5).
    Nakano M., Sugioka K., Ushijima Y., and Goto T.: Chemiluminescence probe with Cypridina luciferin analog, 2-methyl-6-phenyl-3,7-dihydroimidazo[1,2-a]pyrazin-3-one, for estimating the ability of human granulocytes to generate superoxide. Anal. Biochem. 159: 363–369, 1986.CrossRefGoogle Scholar
  6. 6).
    Miyagawa H., Yoshikawa T., Tanigawa T., Yoshida N., Sugino S., Kondo M., Nishikawa H., and Kohno M.: Measurement of serum superoxide dismutase activity by electron spin resonance. J. Clin. Biochem. Nutr., 5: 1–7, 1988.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1990

Authors and Affiliations

  • Toshikazu Yoshikawa
    • 1
  • Yuji Naito
    • 1
  • Toru Tanigawa
    • 1
  • Shigeru Sugino
    • 1
  • Motoharu Kondo
    • 1
  1. 1.First Department of MedicineKyoto Prefectural University of MedicineKamigyo-ku, Kyoto 602Japan

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