Carbon isotope effects associated with Fenton-like degradation of toluene: Potential for differentiation of abiotic and biotic degradation

Hydrogen peroxide (H(2)O(2))-mediated oxygenation to enhance subsurface aerobic biodegradation is a frequently employed remediation technique. However, it may be unclear whether observed organic contaminant mass loss is caused by biodegradation or chemical oxidation via hydroxyl radicals generated during catalyzed Fenton-like reactions. Compound-specific carbon isotope analysis has the potential to discriminate between these processes. Here we report laboratory experiments demonstrating no significant carbon isotope fractionation during Fenton-like hydroxyl radical oxidation of toluene. This implies that observation of significant isotopic fractionation of toluene at a site undergoing H(2)O(2)-mediated remediation would provide direct evidence of biodegradation. We applied this approach at a field site that had undergone 27 months of H(2)O(2)-mediated subsurface oxygenation. Despite substantial decreases (>68%) in groundwater toluene concentrations carbon isotope signatures of toluene (delta(13)C(tol)) showed no significant variation (mean=-27.5+/-0.3 per thousand, n=13) over a range of concentrations from 11.1 to 669.0 mg L(-1). Given that aerobic degradation by ring attack has also been shown to result in no significant isotopic fractionation during degradation, at this site we were unable to discern the mechanism of degradation. However, such differentiation is possible at sites where aerobic degradation by methyl group attack results in significant isotopic fractionation.