title: Toward Streamlined Identification of Dioxin-like Compounds in Environmental Samples through Integration of Suspension Bioassay.
abstract: Effect-directed analysis (EDA) is a powerful strategy to identify biologically active compounds in environmental samples. However, in current EDA studies, fractionation and handling procedures are laborious, consist of multiple evaporation steps, and thus bear the risk of contamination and decreased recoveries of the target compounds. The low resulting throughput has been one of the major bottlenecks of EDA. Here, we propose a high-throughput EDA (HT-EDA) work-flow combining reversed phase high-performance liquid chromatography fractionation of samples into 96-well microplates, followed by toxicity assessment in the micro-EROD bioassay with the wild-type rat hepatoma H4IIE cells, and chemical analysis of bioactive fractions. The approach was evaluated using single substances, binary mixtures, and extracts of sediment samples collected at the Three Gorges Reservoir, Yangtze River, China, as well as the rivers Rhine and Elbe, Germany. Selected bioactive fractions were analyzed by highly sensitive gas chromatography-atmospheric pressure laser ionization-time-of-flight-mass spectrometry. In addition, we optimized the work-flow by seeding previously adapted suspension-cultured H4IIE cells directly into the microplate used for fractionation, which makes any transfers of fractionated samples unnecessary. The proposed HT-EDA work-flow simplifies the procedure for wider application in ecotoxicology and environmental routine programs.
authors.forename: Hongxia
authors.identifier: 0000-0003-0692-4901
authors.initials: H
authors.lastname: Xiao
authors.affiliation: Toxicology Centre and School of Environment and Sustainability, University of Saskatchewan , Saskatoon, Saskatchewan S7N 5B3, Canada.
authors.forename: Markus
authors.initials: M
authors.lastname: Brinkmann
authors.forename: Beat
authors.initials: B
authors.lastname: Thalmann
authors.forename: Andreas
authors.initials: A
authors.lastname: Schiwy
authors.affiliation: Institute of Geology and Palaeontology-Applied Geology, University of Münster , 48149 Münster, Germany.
authors.forename: Sigrid
authors.initials: S
authors.lastname: Große Brinkhaus
authors.affiliation: Institute of Geology and Palaeontology-Applied Geology, University of Münster , 48149 Münster, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Achten
authors.forename: Kathrin
authors.initials: K
authors.lastname: Eichbaum
authors.forename: Carolin
authors.initials: C
authors.lastname: Gembé
authors.forename: Thomas-Benjamin
authors.initials: TB
authors.lastname: Seiler
authors.affiliation: College of Resources and Environmental Science, Chongqing University , 400030 Chongqing, China.|State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University , 210023 Nanjing, China.|College of Environmental Science and Engineering and State Key Laboratory of Pollution Control and Resource Reuse, Tongji University , 200092 Shanghai, China.
authors.forename: Henner
authors.initials: H
authors.lastname: Hollert
mesh_terms: D000818:Animals; D001681:Biological Assay; D004147:Dioxins; D054750:Ecotoxicology; D019015:Geologic Sediments / Q000737:chemistry*; D045483:Rivers / Q000737:chemistry; D014874:Water Pollutants, Chemical
chemical_list: D004147:Dioxins; D014874:Water Pollutants, Chemical