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Development and Application of a Cell-Based Reporter Gene Assay for Detecting (Me-)PAHs in Water Samples

par Su, Yiqi ;Najar, Basma ;Huysmans, Marijke;Van Antwerpen, Pierre ;Gao, Yue;Elskens, Marc
Référence Water Quality, Exposure and Health
Publication Publié, 2025-06-01

Article révisé par les pairs

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Résumé :

Polycyclic aromatic hydrocarbons (PAHs) and their methylated derivatives (Me-PAHs) are (semi-)persistent environmental pollutants with significant toxicological risks. While targeted and non-targeted analyses are effective at identifying individual compounds, they fail to assess the combined biological effects of complex mixtures. This study developed an H1L7.5c1 cell line-based reporter gene assay. Key parameters such as cell density, exposure time, and exposure medium were optimized to enhance the assay sensitivity, precision, and robustness. Our bioassay achieved a limit of detection (LOD) of 0.13-nM benzo[a]pyrene (BaP) equivalents and an effective concentration (EC50) of 1.1 ± 0.1 nM BaP. A relative potency (REP) library was built for 16 PAHs and 5 Me-PAHs. Application to groundwater and wastewater effluent revealed correlations between bioassay-derived and chemically-derived bioequivalent quantity. A close match was observed in groundwater, but more complex wastewater effluent matrices suggested the presence of additional AhR-active compounds. A mixture analysis using a Generalized Response Addition model revealed no significant synergistic or antagonistic interactions among the sample mixtures. This study highlights the potential of using H1L7.5c1 cell line-based bioassay as a rapid and sensitive screening tool for (Me-)PAHs monitoring in environmental samples. It offers a valuable tool for Effect-Based Monitoring and assessment under the EU Water Framework Directive.