Analyzing a broader spectrum of endocrine active organic contaminants in sewage sludge with high resolution LC-QTOF-MS suspect screening and QSAR toxicity prediction

Endocrine active contaminants (EACs) in environmental samples can pose a range of toxicological threats to ecosystems, especially through their impacts on reproductive pathways mediated by the estrogen receptor. The physicochemical properties of known organic EACs vary greatly and typically require different sample preparation techniques to identify different classes of compounds. EAC sources are similarly diverse, including both endogenous compounds and anthropogenic chemicals found in personal care products, pharmaceuticals, and their transformation products, which are often disposed of to sewers at their end of use. Looking for EACs in sewage sludge proposes a bottom-up, or end-of-use and treatment approach to discover environmentally relevant EACs, since many EACs accumulate in sludges even after application of robust wastewater treatment processes. This study demonstrates an extraction and analytical method capable of detecting a broad spectrum of known and suspected EACs via High Resolution Liquid Chromatography Quadropole Time-of-Flight Mass Spectrometry (LC-QTOF-MS) suspect screening of fourteen California sewage sludge samples. Spike-recovery experiments were performed using twelve carefully selected surrogates to assess different extraction solvents, sample weights, extraction pH values, procedures for combining extracts with different extraction pH's, and solid phase extraction cartridges. Using LC-QTOF-MS, identifications of several other organic compounds in the samples were made, a goal unachievable with unit resolution mass spectrometry. Suspect screening of California sludge samples discovered 118 compounds including hormones, pharmaceuticals, phosphate flame retardants, recreational drugs, antimicrobials, and pesticides. Additionally, 22 of these identified compounds are predicted to interfere with estrogen receptors or other reproductive/developmental pathways based on the VEGA QSAR toxicity prediction model. 118 endocrine active compounds were identified in 14 sewage sludge samples using a new sample preparation method that extracts a broad range of compounds analyzed by high-resolution LC-QTOF-MS. Library screening matches were run in a predictive model to identify compounds likely to interfere with estrogen or other reproductive signaling.