Identification of retinoic acid receptor agonists in sewage treatment plants.
Identification of retinoic acid receptor agonists in sewage treatment plants.
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DOI：
10.1021/es9000328
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发表时间：
2009-09
影响因子：
11.4
通讯作者：
Huajun Zhen;Xiaoqin Wu;Jianying Hu;Yang Xiao;Min Yang;J. Hirotsuji;J. Nishikawa;T. Nakanishi
中科院分区：
文献类型：
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作者：
Huajun Zhen;Xiaoqin Wu;Jianying Hu;Yang Xiao;Min Yang;J. Hirotsuji;J. Nishikawa;T. Nakanishi
视黄酸受体（RAR）激动剂被推测是北美广泛观察到的青蛙畸形的一个可能原因，尽管对水生环境中的特定RAR激动剂知之甚少。我们确定了特定的RAR egonists在污水处理厂（STPs）和接收河流使用RAR酵母双杂交生物测定。水样经固相萃取柱提取后，依次用正己烷、乙酸乙酯和甲醇洗脱，进行生物测定。在这三种组分中，乙酸乙酯组分显示出最高的RAR激动活性。生物测定衍生的活性，表示为全反式维甲酸（全反式RA）当量（ATRA-EQ）在STP流入物和流出物中分别为10.9 +/- 2.2和1.7 +/- 1.0 ng/L，而在接受STP流出物的两条河流中，ATRA-EQ分别高达7.1和8.3 ng/L。在生物活性测定的指导下，采用高效液相色谱和超高效液相色谱（UPLC）两步分离，从高碑店STP进水和全反式-4-氧代-RA中分离得到两个活性组分（流入物中4.7-10.4 ng/L，流出物中< 0.2-0.9 ng/L）和13-顺式-4-氧代-RA（流入物中为2.3-7.1纳克/升，< 0.4-1.1 ng/L（流出物中）。RA相对于全反式RA在显示RAR α激动活性方面的相对值分别计算为3.87和0.46。
Retinoic acid receptor (RAR) agonists are speculated to be one possible cause for the widely observed frog deformities in North America, although little is known about the specific RAR agonists in aquatic environments. We identified the specific RAR egonists in sewage treatment plants (STPs) and receiving rivers using an RAR yeast two-hybrid bioassay. Water samples were extracted by solid-phase extract cartridges, which were successively eluted by hexane, ethyl acetate, and methanol for bioassay. Among the three fractions, the ethyl acetate fraction showed the highest RAR agonistic activities. The bioassay-derived activity, expressed as all-trans-retinoic acid (all-transRA) equivalents (ATRA-EQ) were 10.9 +/- 2.2 and 1.7 +/- 1.0 ng/L in the STP influents and effluents, respectively, while the ATRA-EQs were as high as 7.1 and 8.3 ng/L in the two rivers receiving STP effluents. Following a two-step fractionation using high-performance liquid chromatography and ultra-performance liquid chromatography (UPLC) directed by the bioassay, two bioactive fractions were obtained from Gaobeidian STP influent and all-trans-4-oxo-RA (4.7-10.4 ng/L in influents, < 0.2-0.9 ng/L in effluents) and 13-cis-4-oxo-RA (2.3-7.1 ng/L in influents, < 0.4-1.1 ng/L in effluents) were identified in these fractions with UPLC-MS/MS. The EC50 for all-trans-4-oxo-RA or 13-cis-4-oxo-RA relative to that of all-trans-RA in exhibiting RARalpha agonistic activity was calculated to be 3.87 and 0.46, respectively.