title: Rapid activity-directed screening of estrogens by parallel coupling of liquid chromatography with a functional gene reporter assay and mass spectrometry.
abstract: In this study we developed a new LC nanofractionation platform that combines a human cell (BG1.Luc) gene reporter assay with a high resolution mass spectrometer for the detection and identification of estrogenic and anti-estrogenic compounds in environmental waters. The selection of this assay was based on its high sensitivity and selectivity, which is required for environmental trace level detection. We modified an autosampler and controlled it with in-house developed software to collect fractions in the low second range in microtiter plates. This ensured that chromatographic separation was maintained and allowed straightforward hyphenation with the bioassay. After bioassay testing, bioassay chromatograms were reconstructed and directly correlated with MS chromatograms that were obtained in parallel. This enabled to pinpoint bioactives in the MS chromatogram within a single fractionation cycle and results in a significant increase in throughput compared to traditional EDA studies. The sensitivity of the platform was low enough for environmental waters (80nM for bisphenol A and 320pM and 3.2nM for estradiol and estriol, respectively). In addition, the ability of the platform to detect anti-estrogens was successfully demonstrated as well. Finally, real samples were analysed.
authors.affiliation: VU University, Division of BioAnalytical Chemistry, de Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
authors.forename: Willem
authors.initials: W
authors.lastname: Jonker
authors.affiliation: VU University, Institute for Environmental Studies, de Boelelaan 1087, 1081 HV Amsterdam, The Netherlands.
authors.forename: Marja H
authors.initials: MH
authors.lastname: Lamoree
authors.affiliation: The Water Laboratory, J.W. Lucasweg 2, 2031 BE Haarlem, The Netherlands.
authors.forename: Corine J
authors.initials: CJ
authors.lastname: Houtman
authors.affiliation: VU University, Institute for Environmental Studies, de Boelelaan 1087, 1081 HV Amsterdam, The Netherlands.
authors.forename: Timo
authors.initials: T
authors.lastname: Hamers
authors.affiliation: VU University, Division of BioAnalytical Chemistry, de Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
authors.forename: Govert W
authors.initials: GW
authors.lastname: Somsen
authors.affiliation: VU University, Division of BioAnalytical Chemistry, de Boelelaan 1083, 1081 HV Amsterdam, The Netherlands. Electronic address: j.kool@vu.nl.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Kool
mesh_terms: D001681:Biological Assay; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation* / Q000379:methods*; D002853:Chromatography, Liquid; D004784:Environmental Monitoring / Q000379:methods*; D004958:Estradiol / Q000032:analysis; D004967:Estrogens / Q000032:analysis*; D013058:Mass Spectrometry; D014867:Water / Q000737:chemistry*; D014874:Water Pollutants, Chemical / Q000032:analysis
chemical_list: D004967:Estrogens; D014874:Water Pollutants, Chemical; D014867:Water; D004958:Estradiol
keywords: Effect-directed analysis; Estrogen; Fractionation; High resolution; High throughput; Toxicant