Identification of emerging pharmaceutical pollutants in a river impacted by an industrial effluent combining passive sampling and effect-directed analysis

Open Access

Abstract

The occurrence of endocrine disrupting compounds was previously detected in a French river impacted by an urban and a pharmaceutical wastewater treatment plants (WWTPs). Indeed, first investigations were conducted in surface water using polar organic compound integrative sampler (POCIS) at upstream and downstream of the WWTPs. Toxicological profiling of POCIS extracts was evaluated using in vitro bioassays based on luciferase gene reporter. Strong activities were found in POCIS extracts downstream from pharmaceutical factory. The identification of 60 chemicals (pharmaceuticals, antibiotics and steroids) by targeted chemical analyses (LC-MS/MS) did not totally explain the activities. In order to isolate the active chemicals and to identify them, POCIS crude extract was fractionated using Reverse Phase – High Performance Liquid Chromatography (RP-HPLC). Each fraction was then tested individually on in vitro cell lines. Several steroid compounds were detected by target analyses using LC-MS/MS and these chemicals explained the majority of observed activities in the selected fractions. For many highly active fractions, compounds responsible for these activities remained unknown. In this study, a LC-HRMS system (LC-QTOF) was used to identify these active compounds. The HPLC fractions were analyzed in (data-dependent) auto-MS/MS acquisition mode. Detected compounds in fractions of POCIS extract were compared to those found in the corresponding fractions of the blank sample. The characterization of compounds was performed on molecules only present in POCIS fractions. The procedure allowed the identification of several candidate molecules including drugs, steroids and some of their metabolites. The identification of some of them has been confirmed with purchased reference standards. The activity of these identified chemicals is still under process and results will be presented. Acknowledgements: The French Ministry of Ecology, Sustainable Development and Energy, the Aquitaine Region and the European Union (CPER A2E project) are acknowledged for their financial support. Europe is moving in Aquitaine with the European Regional Development Fund.