Synthesis of peroxynitrite from nitrite and hydrogen peroxide.
We report a simple method for the synthesis of peroxynitrite from nitrite and hydrogen peroxide that can generate hundreds of milliliters of 180 mM peroxynitrite within 1 h from start to finish. It can be scaled down to make small quantities of isotope-labeled peroxynitrite. The method requires only a syringe pump and tubing connectors and is feasible for any biochemical laboratory. Unreacted hydrogen peroxide is eliminated with manganese dioxide, using an improved preparation compared to commercially available manganese dioxide. A number of contaminants were detected by mass spectrometry in peroxynitrite solutions cleaned with commercially purchased manganese dioxide. Nitrite contamination of the peroxynitrite solution is less than 2% as determined using the Griess method. The residual contaminants are principally 0.28 M sodium chloride and 0.1 M sodium hydroxide, which pose few problems when peroxynitrite is diluted for use in biological experiments.
Robinson
Kristine M
KM
Department of Biochemistry and Biophysics, Oregon State University, Environmental Health Science Center, The Linus Pauling Institute Corvallis, OR 97331, USA.
Beckman
Joseph S
JS
D006861:Hydrogen Peroxide / Q000737:chemistry*; D017895:Manganese Compounds / Q000737:chemistry; D013058:Mass Spectrometry; D009573:Nitrites / Q000737:chemistry*; D010087:Oxides / Q000737:chemistry; D030421:Peroxynitrous Acid / Q000138:chemical synthesis*
D017895:Manganese Compounds; D009573:Nitrites; D010087:Oxides; D030421:Peroxynitrous Acid; D006861:Hydrogen Peroxide; C016552:manganese dioxide