title: Diazotization reaction of nitric oxide trapped by hemoglobin.
abstract: The present study addresses to determine whether hemoglobin within red blood cells can be utilized as a spin-trap agent for nitric oxide. We demonstrate the diazotization method coupled with a gel filtration chromatography, which is simply due to the separation of nitrosylhemoglobin from nitrite, nitrate or other low molecular nitroso-compounds in biological systems and to the liberation of nitric oxide from nitrosyl heme-complexes in the acidic condition. The amount of nitric oxide can be estimated by the difference of absorbances at 542 nm between diazo-compounds formed by Griess reagent and hemichrome by phosphoric acid. Our results indicate that hemoglobin in red cells as a spin-trap agent would be useful for monitoring nitric oxide in the circulation under the several disease states.
authors.affiliation: 1st. Department of Biochemistry, Saitama Medical School, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Sonoda
authors.forename: T
authors.initials: T
authors.lastname: Hashimoto
authors.forename: A
authors.initials: A
authors.lastname: Satomi
authors.forename: T
authors.initials: T
authors.lastname: Miyazaki
authors.forename: K
authors.initials: K
authors.lastname: Ishida
authors.forename: Y
authors.initials: Y
authors.lastname: Sakagishi
mesh_terms: D002850:Chromatography, Gel; D003979:Diazonium Compounds; D004578:Electron Spin Resonance Spectroscopy; D004912:Erythrocytes / Q000378:metabolism; D006454:Hemoglobins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008991:Monitoring, Physiologic; D009569:Nitric Oxide / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D013053:Spectrophotometry
chemical_list: D003979:Diazonium Compounds; D006454:Hemoglobins; C008657:nitrosyl hemoglobin; D009569:Nitric Oxide