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2003 Fiscal Year Final Research Report Summary

Development for measuring the concentration of hydrogen peroxide in vivo using ESR equipment

Research Project

Project/Area Number 14572044
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Physical pharmacy
Research InstitutionNational Institute of Radiological Sciences

Principal Investigator

UEDA Jun-ichi  National Institute of Radiological Sciences, Redox Regulation Research Group, Team Leader, 放射線安全研究センター・レドックス制御研究グループ, チームリーダー (90160168)

Co-Investigator(Kenkyū-buntansha) TAKESHITA Keizo  National Institute of Radiological Sciences, Redox Regulation Research Group, Senior Researcher, 放射線安全研究センター・レドックス制御研究グループ, 主任研究員 (70175438)
Project Period (FY) 2002 – 2003
Keywordshydrogen peroxide / free radical / in vivo ESR / in vitro ESR / L-band / surface-coil-type resonator
Research Abstract

In order to measure quantitatively the amount of hydrogen peroxide (H_2O_2) generated by phagocytes or irradiation of ionizing radiation in whole body, a cation radical formed by the reaction of H_2O_2 with 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) in the presence of peroxidase (HRP) was measured using L-band ESR with surface-coil-type resonator. The amount of H_2O_2 may be related to signal intensity of the ESR spectrum of ABTS cation radical, where H_2O_2 reacts with HRP, resulting in the production of compound I, and then the compound I oxidized ABTS to yield ABTS cation radical.
Liposomes (1.24 mol ABTS/mol phospholipid (DMPC : cholesterol : stearylamine=1:1:0.1)) containing ABTS and HRP (ABTS-HRP-liposome) were prepared. However, the liposomes were not used for further analysis.
A solution of pre-formed ABTS cation radical solution produced by mixing an ABTS-HRP solution with various concentrations of H_2O_2,was intraperitoneally injected into mouse and measured using a L-band ESR spectrometer to elucidate sensitivity of the L-band ESR. The L-band ESR signal of ABTS cation radical was observed in the presence of H_2O_2 more than 2 ? mol, suggesting that the lowest limiting concentration of H_2O_2 is probably 2 ? mol.
On the other hand, the ABTS-HRP solution was intraperitoneally injected into mouse, followed by the injection of a large amount of H_2O_2 into mouse. As a result, the L-band ESR spectrum of ABTS cation radical was obtained. On the contrary, the injection of LPS, which should cause the generation accumulation of phagocytes in the peritoneal cavity, gave no ESR spectrum.

  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] Keita Saito: "Two reaction sites of a spin label, TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl), with hydroxyl radical"J.Pharm.Sci.. 92. 1-6 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Jun-ichi Ueda: "Singlet oxygen-mediated hydroxyl radical production in the presence of phenols : Whether DMPO-OH formation really indicates production of OH?"Photochem.Photobiol.. 77. 165-170 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Akira Hanaki: "Transport of the Cu(II) bound with histidine-containing tripeptides to cysteine. Coordination mode and exchangeability of Cu(II) in the complexes."Bull.Chem.Soc.Jpn.. 76. 2143-2150 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Chiho Nishizawa: "Hydroxyl radical generation caused by the reaction of singlet oxygen with a spin trap, DMPO, increases significantly in the presence of biological reductants"Free Radic.Res.. 38. 385-392 (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Jun-ichi Ueda: "Reactive oxygen species scavenging ability of a new compound derived from weathered coal"Spectrochimica Acta Part A. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Saito et al.: "Two reaction sites of a spin label, TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl), with hydroxyl radical"J.Pharm.Sci.. 92. 1-6 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Ueda et al.: "Singlet oxygen-mediated hydroxyl radical production in the presence of phenols : Whether DMPO-OH formation really indicates production of OH?"Photochem.Photobiol.. 77. 165-170 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A.Hanaki et al.: "Transport of the Cu(II) bound with histidine-containing tripeptides to cysteine. Coordination mode and exchangeability of Cu(II) in the complexes"Bull.Chem.Soc.Jpn.. 76. 2143-2150 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] C.Nishizawa et al.: "Hydroxyl radical generation caused by the reaction of singlet oxygen with a spin trap, DMPO, increases significantly in the presence of biological reductants"Free Radic.Res.. 38. 385-392 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Ueda et al.: "Reactive oxygen species scavenging ability of a new Compound derived from weathered coal"Spectrochimica Acta Part A. (in press).

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2005-04-19  

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