Hydroxyl radical formation upon oxidation of reduced humic acids by oxygen in the dark

Humic acids (HAs) accept and donate electrons in many biogeochemical redox reactions at oxic/anoxic interfaces. The products of oxidation of reduced HAs by O ₂ are unknown but are expected to yield reactive oxygen species, potentially including hydroxyl radical (·OH). To quantify the formation of ·OH upon oxidation of reduced HAs by O ₂, three HAs were reduced electrochemically to well-defined redox states and were subsequently oxidized by O ₂ in the presence of the ·OH probe terephthalate. The formation of ·OH upon oxidation increased with increasing extent of HA reduction. The yield of ·OH ranged from 42 to 160 mmol per mole of electrons donated by the reduced HA. The intermediacy of hydrogen peroxide (H ₂O ₂) in the formation of ·OH was supported by enhancement of ·OH formation upon addition of exogenous H ₂O ₂ sources and by the suppression of ·OH formation upon addition of catalase as a quencher of endogenous H ₂O ₂. The formation of ·OH in the dark during oxidation of reduced HA represents a previously unknown source of ·OH formation at oxic/anoxic interfaces and may affect the biogeochemical and pollutant redox dynamics at these interfaces.