Toxicity of Ozonated Wastewater to HepG2 Cells: Taking Full Account of Nonvolatile, Volatile, and Inorganic Byproducts.

Wastewater ozonation forms various toxic byproducts, such as aldehydes, bromate, and organic bromine. However, there is currently no clear understanding of the overall toxicity changes in ozonated wastewater because pretreatment with solid phase extraction cannot retain inorganic bromate and volatile aldehydes, yet contributions of known ozonation byproducts to toxicity are unknown. Moreover, compared with bromate, organic bromine did not receive widespread attention. This study evaluated the toxicity of ozonated wastewater by taking aldehydes, bromate, and organic bromine into consideration. In the absence of bromide, formaldehyde contributed 96-97% cytotoxicity and 92-95% genotoxicity to HepG2 cells among the detected known byproducts, while acetaldehyde, propionaldehyde, and glyoxal had little toxicity. Both formaldehyde and dibromoacetonitrile drove toxicity among the known byproducts when bromide was present. Toxicity assays in HepG2 cells showed that when secondary effluents contained no bromide, the cytotoxicity of the nonvolatile organic fraction (NVOF) was reduced by 56-70%, and genotoxicity was completely removed after ozonation. However, the formed aldehydes (volatile organic fraction, VOF) led to increased overall toxicity. In the presence of bromide, compared with the secondary effluent, ozonation increased the cytotoxicity of the NVOFBr from 3.4-4.0 mg phenol/L to 10.3-13.9 mg phenol/L, possibly due to the formation of organic bromine. In addition, considering the toxicity of VOFBr (VOF in the presence of bromide, including aldehydes, tribromomethane, etc.), the overall cytotoxicity and genotoxicity became much higher than those of the secondary effluent. Although bromate had a limited impact on cytotoxicity and genotoxicity, it caused an increase in oxidative stress in HepG2 cells. Therefore, when taking full account of nonvolatile, volatile, and inorganic fractions, ozonation generally increases the toxicity of wastewater.