Effect-directed analysis of endocrine-disrupting compounds in multi-contaminated sediment: identification of novel ligands of estrogen and pregnane X receptors
Effect-directed analysis of endocrine-disrupting compounds in multi-contaminated sediment: identification of novel ligands of estrogen and pregnane X receptors
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DOI：
10.1007/s00216-013-6708-5
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发表时间：
2013-03-01
影响因子：
4.3
通讯作者：
Ait-Aissa, Selim
中科院分区：
文献类型：
--
作者：
Creusot, Nicolas;Budzinski, Helene;Ait-Aissa, Selim
越来越多地采用基于环境导向分析的战略，以确定不利（生态）毒性效应与化学污染物之间的因果关系。在这项研究中，我们报告的开发和使用的EDA方法来确定内分泌干扰化学品（EDCs）在多污染的河流沉积物。电池的体外报告细胞为基础的生物测定，测量雌激素，（抗）雄激素，二恶英样，和X-ray烷X受体（PXR）样的活动，揭示了多污染的配置文件。为了分离宽极性范围的活性化合物，我们建立了一个多步分馏程序，该程序结合：（1）使用基于正相的固相萃取（SPE）的初级分馏步骤，用12种非极性至极性标准EDCs的混合物进行验证;（2）使用基于反相的高效液相色谱（RP-HPLC）的二级分馏，用33种标准EDCs校准;和（3）使用重组雌激素受体（ER）亲和柱的纯化步骤。体外SPE和HPLC分析表明，ER和PXR活性主要是由于极性到中等极性的化合物，而二恶英样和抗雄激素活性在极性较低的馏分。整个过程允许最终分离和鉴定新的环境PXR（例如，邻苯二甲酸二异辛酯）和ER（例如，2，4-二叔丁基苯酚和2，6-二叔丁基-α-甲氧基-对甲酚）配体。使用商业标准进一步证实了这些化学品的体外生物活性，其中邻苯二甲酸二异辛酯首次被鉴定为有效的hPXR环境激动剂。
Effect-directed analysis (EDA)-based strategies have been increasingly used in order to identify the causative link between adverse (eco-)toxic effects and chemical contaminants. In this study, we report the development and use of an EDA approach to identify endocrine-disrupting chemicals (EDCs) in a multi-contaminated river sediment. The battery of in vitro reporter cell-based bioassays, measuring estrogenic, (anti)androgenic, dioxin-like, and pregnane X receptor (PXR)-like activities, revealed multi-contamination profiles. To isolate active compounds of a wide polarity range, we established a multi-step fractionation procedure combining: (1) a primary fractionation step using normal phase-based solid-phase extraction (SPE), validated with a mixture of 12 non-polar to polar standard EDCs; (2) a secondary fractionation using reversed-phase-based high-performance liquid chromatography (RP-HPLC) calibrated with 33 standard EDCs; and (3) a purification step using a recombinant estrogen receptor (ER) affinity column. In vitro SPE and HPLC profiles revealed that ER and PXR activities were mainly due to polar to mid-polar compounds, while dioxin-like and anti-androgenic activities were in the less polar fractions. The overall procedure allowed final isolation and identification of new environmental PXR (e.g., di-iso-octylphthalate) and ER (e.g., 2,4-di-tert-butylphenol and 2,6-di-tert-butyl-alpha-methoxy-p-cresol) ligands by using gas chromatography coupled with mass spectrometry with full-scan mode acquisition in mid-polar fractions. In vitro biological activity of these chemicals was further confirmed using commercial standards, with di-iso-octylphthalate identified for the first time as a potent hPXR environmental agonist.