Oxygen-accepting antioxidants which arise during ascorbate oxidation

To identify potential antioxidant compounds derived from ascorbate, the hydrogen peroxide-induced oxidation of ascorbate and dehydroascorbate was studied by gas-chromatography electron impact mass spectrometry and liquid-chromatography electrospray mass spectrometry in real time. Significant differences in ascorbate and dehydroascorbate oxidation occurred at pH 3.3 compared to pH 7.4. Of note, the primary species present in dehydroascorbate (DHA)-containing solutions at pH 7.4 had a spectrum consistent with diketogulonate. Hydrogen peroxide exposure of DHA-containing solutions formed threonate more rapidly at pH 7.4 than at pH 3.3. In these solutions, a 5-carbon species with mass spectral characteristics of a 3,4,5-trihydroxy-2-ketopentanoate appeared to be an intermediate between diketogulonate and threonate, and was more labile than other species in the presence of hydrogen peroxide. These data suggest that a 3,4,5-trihydroxy-2-ketopentanoate is potentially a key antioxidant compound in the ascorbate degradation cascade and in ascorbate-containing solutions at physiologic pH.