Mechanism of initial reaction of phenylhydrazine with oxyhemoglobin and effect of ring substitutions on the biomolecular rate constant of this reaction.

ABSTRACT

Phenylhydrazine in the presence of oxygen causes the oxidative denaturation of hemoglobin. The initial step in this process is a bimolecular reaction, probably a two-electron transfer from phenylhydrazine to oxyhemoglobin. The product of this reaction is neither methemoglobin nor deoxyhemoglobin. Superoxide dismutase and catalase eliminate side reactions that increase the apparent rate of this reaction as measured spectrophotometrically at 577 nm; scavengers for the hydroxyl radical and singlet oxygen do not affect this rate either in the presence or in the absence of these enzymes. Halogen atoms and alkyl groups decrease the rate when ortho and increase the rate when meta or para to the hydrazino group. Chlorine atoms at both ortho positions or the carboxylate group at the ortho or the para position block the reaction. In the presence of phenylhydrazine under air, methemoglobin is converted to the same complex as that produced when phenyldiazene is added to methemoglobin anaerobically. Under N2 or CO, phenylhydrazine reduces methemoglobin to deoxyhemoglobin or carbonmonoxyhemoglobin.