Biological Validation of a Sample Preparation Method for ER-CALUX Bioanalysis of Estrogenic Activity in Sediment Using Mixtures of Xeno-Estrogens
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- 作者：Corine J. Houtman ; Yoni K. van Houten ; Pim E. G. Leonards ; Abraham Brouwer ; Marja H. Lamoree ; and Juliette Legler
- 刊名：Environmental Science & Technology
- 出版年：2006
- 出版时间：April 1, 2006
- 年：2006
- 卷：40
- 期：7
- 页码：2455 - 2461
- 全文大小：146K
- 年卷期：v.40,no.7(April 1, 2006)
- ISSN：1520-5851
文摘
The combined estrogenic effects of mixtures of environmental pollutants in the in vitro ER-CALUX (chemicalactivated luciferase gene expression) bioassay were examinedto biologically validate a sample preparation method forthe analysis of estrogenic compounds in sediment. The methodused accelerated solvent extraction (ASE) and gelpermeation chromatography (GPC) and was validated withrespect to recovery of biological response taking mixtureeffects 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. Experimentallydetermined mixture effects were well described by theconcept of concentration addition (CA), as expected forsimilarly acting compounds. Observed estradiol equivalencefactors of the mixtures (on average 1.2 ± 0.3) agreedvery well with the value predicted according to CA. Thesample preparation method was then applied to pure mixturesof standards and to sediment spiked with one of themixtures. Recoveries of estrogenic compounds wereestimated by determination of their mixture potencies in ER-CALUX and compared to the mixture effects predictedby CA. Recoveries of estrogenic activity were between 80and 129%, indicating that the additive behavior of mixturesof xeno-estrogens is well conserved during samplepreparation. Together with an average repeatability of18.3%, low average limit of detection (2.6 ± 1.8 pg of EEQ/g), and coefficient of variance (3.5 ± 3.3%), this demonstratedthe suitability of the sample preparation method for theanalysis of mixtures of (xeno-)estrogenic compounds insediment with the ER-CALUX assay.
-chlordane,dibutylphthalate, (4,4'-dichlorodiphenyl)dichloroethylene,and 2,4,5-trichlorobiphenyl) were prepared. Experimentallydetermined mixture effects were well described by theconcept of concentration addition (CA), as expected forsimilarly acting compounds. Observed estradiol equivalencefactors of the mixtures (on average 1.2 ± 0.3) agreedvery well with the value predicted according to CA. Thesample preparation method was then applied to pure mixturesof standards and to sediment spiked with one of themixtures. Recoveries of estrogenic compounds wereestimated by determination of their mixture potencies in ER-CALUX and compared to the mixture effects predictedby CA. Recoveries of estrogenic activity were between 80and 129%, indicating that the additive behavior of mixturesof xeno-estrogens is well conserved during samplepreparation. Together with an average repeatability of18.3%, low average limit of detection (2.6 ± 1.8 pg of EEQ/g), and coefficient of variance (3.5 ± 3.3%), this demonstratedthe suitability of the sample preparation method for theanalysis of mixtures of (xeno-)estrogenic compounds insediment with the ER-CALUX assay.