Effect-Based High-Throughput Screening of Environmental Water Samples: Focus on Liver Toxicity Pathways and Endocrine Disruption

The rapid expansion of industrial activities has led to the increased release of harmful pollutants into groundwater, posing significant risks to human health and the environment. Traditional toxicity assessments, which rely on animal testing or limited chemical analyses, often fail to capture complex biological effects or efficiently prioritize risks. To address these challenges, we developed an effect-based framework that integrates high-throughput screening (HTS), high-content screening (HCS), Adverse Outcome Pathways (AOPs), and computational ToxPi analysis. We evaluated the framework using four common industrial contaminants and applied it to groundwater samples from a petrochemical plant. Cell-based assays identified the samples with potential liver and kidney toxicity, evidenced by high cytotoxicity, increased mitochondrial reactive oxygen species (ROS) production, apoptosis, and endocrine-disrupting effects, such as altered steroid hormone production. ToxPi analysis integrated the results from multiple endpoints to generate composite scores and enabled the systematic ranking of groundwater samples according to their overall potential risk. Our findings demonstrate that combining HTS/HCS assays with mechanistic insights from AOPs and computational analysis provides a comprehensive and efficient strategy for groundwater toxicity screening, supporting more effective environmental monitoring and public health protection.