Identifying and prioritizing organic toxicants in treated flowback and produced water from shale gas exploitation sites using an integrative effect-directed analysis and nontarget screening method

The use of hydraulic fracturing in shale gas exploitation has generated substantial amount of flowback and produced water (FPW), and ecological risk of these highly complex chemical mixtures has raised worldwide concern. Herein, an integrative effect-directed analysis (EDA) and nontarget screening (NTS) workflow was developed to identify and prioritize main toxicants in the treated FPW (T-FPW). The workflow included sample extraction and fractionation, zebrafish embryo toxicity tests, target and nontarget chemical analyses, and toxicity prioritization and confirmation using toxicological priority index (ToxPi). Results showed that less hydrophobic compounds (log K-ow < 3.7) which were used in fracturing fluid and their degradation products were the potentially high-risk toxicants in T-FPW. Thirty-nine target compounds identified in toxic fraction explained 4.82% of the mortality. Additional 584 nontarget contaminants were annotated by NTS. Risk prioritization was achieved for 470 identified contaminants with ecotoxicity data available using a ToxPi method. Six nontarget toxicants were identified with higher ecological risks than all target contaminants, and their presence in FPW were confirmed using reference standards. A principal component analysis of NTS features revealed that EDA fractionation reduced mixture complexity and focused toxicant screening, which significantly improved NTS efficiency, highlighting advantages of integrative EDA and NTS for mixture risk assessment.