Diazotization reaction of nitric oxide trapped by hemoglobin.
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.
Sonoda
M
M
1st. Department of Biochemistry, Saitama Medical School, Japan.
Hashimoto
T
T
Satomi
A
A
Miyazaki
T
T
Ishida
K
K
Sakagishi
Y
Y
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
D003979:Diazonium Compounds; D006454:Hemoglobins; C008657:nitrosyl hemoglobin; D009569:Nitric Oxide