Hydrogen Peroxide → Hydrogen Peroxide

Participants

Reaction Steps

Peroxynitrite was prepared as described previously (17). Briefly, an aqueous solution of 0.6 M sodium nitrite was rapidly mixed with an equal volume of 0.7 M hydrogen peroxide containing 0.6 M HCl and immediately quenched with the same volume of 1.5 M NaOH. All reaction solutions were kept on ice. The concentration of peroxynitrite was determined spectrally in 0.1 M NaOH (ε302 nm=1,670 M−1cm−1) (18). The concentration of freshly synthesized peroxynitrite solutions was typically 150 mM 180 mM (75% of theoretical yield) leaving a 25% contamination (50 mM) by of H2O2 and nitrite. Thus, when 20 μM peroxynitrite was used to oxidize Mn porphyrins, 5 μM each of H2O2 and nitrite were present. Tetramethylammonium peroxynitrite (purchased from Alexis), which has no H2O2 contamination, was used to assess any interference due to H2O2. H2O2 increased the rate of re-reduction of Mn(IV)porphyrins back to their respective Mn(III) forms, apparently via direct reduction of the Mn(IV)=O intermediate. However, the rate of H2O2-mediated re-reduction was roughly 1500-fold slower than oxidation of Mn(III)porphyrins by peroxynitrite. Thus, interference with peroxynitrite-mediated metalloporphyrin oxidation by the low micromolar amounts of H2O2 present was negligible.

Provenance & Evidence