Abstract
Wastewater ozonation is commonly employed to enhance the subsequent biodegradation of effluent organic matter (EfOM) and contaminants of concern. However, there is evidence suggesting the formation of recalcitrant ozonation products (OPs) from EfOM. To investigate the biodegradability of OPs we conducted batch biodegradation experiments using wastewater effluent ozonated with mass-labeled ((18)O) ozone. Molecular level analysis of EfOM was performed using reversed-phase liquid chromatography coupled with Fourier transform ion cyclotron resonance mass spectrometry (LC-FT-ICR MS) after 3 and 28 days in batch bioreactors. The use of mass labeling allowed for the unambiguous detection of OPs, with 2933 labeled OP features identified in the ozonated EfOM. Furthermore, employing polarity separation with LC facilitated the investigation of reactivity among different OP isomers. Overall, OPs exhibited a comparable proportion of recalcitrant and bioproduced molecular formulas when compared to the remaining EfOM, with 12% of OPs classified as recalcitrant and 17% bioproduced, indicating that OPs themselves are not inherently biodegradable. Additionally, recalcitrant OPs were found to be more polar based on the O/C ratios and retention time, in comparison to biodegradable OPs. Approximately one-third of the OP isomers displayed variations in their biodegradability, suggesting that studying the degradability of ozonated EfOM at the molecular formula level is heavily influenced by structural differences. Here, we offer unique insight into the biological transformation of EfOM after ozonation using labeled ozone and LC-FT-ICR MS analysis.