- Y.K. Houten Van (1)
- C.J. Houtman (2)
- H.J.C. Klamer (1)
- M.H. Lamoree (3)
- J. Legler (2)
- P.E.G. Leonards (2)
- A.J. Murk (1)
- L.A. Villerius (1)
Biolonical validation of a sample preparation method for ER-CALUX bioanalysis of estrogenic activity in sediment using mixtures of xeno-estrogens
Houtman, C.J. ; Houten, Y.K. Van; Leonards, P.E.G. ; Brouwer, A. ; Lamoree, M.H. ; Legler, J. - \ 2006
Environmental Science and Technology 40 (2006)7. - ISSN 0013-936X - p. 2455 - 2461.
in-vitro bioassays - cell-proliferation - xenoestrogens - chemicals - assay
The combined estrogenic effects of mixtures of environmental pollutants in the in vitro ER-CALUX (chemical activated luciferase gene expression) bioassay were examined to biologically validate a sample preparation method for the analysis of estrogenic compounds in sediment. The method used accelerated solvent extraction (ASE) and gel permeation chromatography (GPC) and was validated with respect to recovery of biological response taking mixture effects into account. Four mixtures of three to six xeno-estrogenic compounds (bisphenol A, 4-nonylphenol, (4,4`-dichlorodiphenyl)trichloroethane, (2,4`-dichlorodiphenyl)trichloroethane, dieldrin, 4-n-octylphenol, ¿-chlordane, dibutylphthalate, (4,4`-dichlorodiphenyl)dichloroethylene, and 2,4,5-trichlorobiphenyl) were prepared. Experimentally determined mixture effects were well described by the concept of concentration addition (CA), as expected for similarly acting compounds. Observed estradiol equivalence factors of the mixtures (on average 1.2 ± 0.3) agreed very well with the value predicted according to CA. The sample preparation method was then applied to pure mixtures of standards and to sediment spiked with one of the mixtures. Recoveries of estrogenic compounds were estimated by determination of their mixture potencies in ER-CALUX and compared to the mixture effects predicted by CA. Recoveries of estrogenic activity were between 80 and 129%, indicating that the additive behavior of mixtures of xeno-estrogens is well conserved during sample preparation. Together with an average repeatability of 18.3%, low average limit of detection (2.6 ± 1.8 pg of EEQ/g), and coefficient of variance (3.5 ± 3.3%), this demonstrated the suitability of the sample preparation method for the analysis of mixtures of (xeno-)estrogenic compounds in sediment with the ER-CALUX assay
A chemical and toxicological profile of Dutch North Sea surface sediments
Klamer, H.J.C. ; Leonards, P.E.G. ; Lamoree, M.H. ; Villerius, L.A. ; Akerman, J.E. ; Bakker, J.F. - \ 2005
Chemosphere 58 (2005)11. - ISSN 0045-6535 - p. 1579 - 1587.
polycyclic aromatic-hydrocarbons - polybrominated diphenyl ethers - flame retardants - coastal waters - irgarol-1051 - extracts - biota - river - assay
Chemical and toxicological profiles were assessed in surface sediments (fraction
Toxicological profiling of sediments with in vitro mechanisms-based bioassays for endocrine disruption
Houtman, C.J. ; Cenijn, P.H. ; Hamers, T. ; Lamoree, M.H. ; Legler, J. ; Murk, A.J. ; Brouwer, A. - \ 2004
Environmental Toxicology and Chemistry 23 (2004)1. - ISSN 0730-7268 - p. 32 - 40.
biotesten - sediment - toxiciteit - hormonen - estuaria - rivieren - nederland - hormoonverstoorders - waterbodems - ecotoxicologie - rijn - maas - bioassays - sediment - toxicity - hormones - estuaries - rivers - netherlands - endocrine disruptors - water bottoms - ecotoxicology - river rhine - river meuse - reporter gene assays - estrogenic activity - aromatic-hydrocarbons - human transthyretin - expression assays - toxic potency - extracts - chemicals - exposure - wildlife
In vitro bioassays are valuable tools for screening environmental samples for the presence of bioactive (e.g., endocrine-disrupting) compounds. They can be used to direct chemical analysis of active compounds in toxicity identification and evaluation (TIE) approaches. In the present study, five in vitro bioassays were used to profile toxic potencies in sediments, with emphasis on endocrine disruption. Nonpolar total and acid-treated stable extracts of sediments from 15 locations in the Rhine Meuse estuary area in The Netherlands were assessed. Dioxin-like and estrogenic activities (using dioxin-responsive chemical-activated luciferase gene expression [DR-CALUX] and estrogen-responsive chemical-activated luciferase gene expression [ER-CALUX] assays) as well as genotoxicity (UMU test) and nonspecific toxic potency (Vibrio fischeri assay) were observed in sediment extracts. For the first time, to our knowledge, in vitro displacement of thyroid hormone thyroxine (T4) from the thyroid hormone transport protein thransthyretin by sediment extracts was observed, indicating the presence of compounds potentially able to disrupt T4 plasma transport processes. Antiestrogenic activity was also observed in sediment. The present study showed the occurrence of endocrine-disrupting potencies in sediments from the Dutch delta and the suitability of the ER- and DR-CALUX bioassays to direct endocrine-disruption TIE studies.