臺灣博碩士論文加值系統

內分泌干擾物(Endocrine Disrupting Chemicals, EDCs)是近年來熱門的研究議題，這類化合物存在於各式工業與生活用品中，並會藉由皮膚接觸、吸入、攝入或是母體暴露等途徑進入到人體，除了暴露途徑廣泛外其在非常低的濃度下即可產生內分泌的干擾如發育障礙、生殖損傷、肥胖、癌症等疾病，其中EDCs中天然與合成的雌激素會經由人體或動物排洩而排放至環境中，更有一類化合物因化學結構、受體特徵及信號傳導類似雌激素又稱為外源性雌激素(Xenoestrogens, XEs)。
武洛溪主要汙水來源為畜牧廢水及生活汙水，由於沿岸有高密度動物畜牧場，故其天然排泄之雌激素值得關注。本研究針對武洛溪流域上游至下游12個樣站點進行為期4季之採樣調查其水質狀況，並利用生物分析法T47D-Kbluc冷光報導基因法與E-Screen細胞增生法，配合HPLC-MS2儀器分析來評估武洛溪沿岸之雌激素濃度與活性分布。根據四季採樣結果武洛溪水質RPI評估後水體狀況為中度汙染至嚴重汙染。
而根據生物試驗結果顯示武洛溪流域均有高雌激素活性存在，並隨著乾溼季不同而有所變化，T47D-Kbluc原水樣品活性範圍介於0.23~5.62 ng/L E2T47D-KblucEEQ，而T47D-Kbluc濃縮樣品活性範圍介於11.66~190.06 ng/L E2T47D-KblucEEQ；E-Screen細胞增生法之濃縮樣品活性範圍介於6.63~84.50 ng/L E2E-ScreenEEQ。根據化學分析之結果顯示，檢出率高於85%的化合物有雌酮(Estrone, E1)、雙酚A(Bisphenol A, BPA)、壬基苯酚雙乙氧基醇(Nonylphenol di-ethoxylate, NP2EO)、二苯甲酮(Benzophenone, BP)、二苯甲酮-3(Benzophenone-3, BP-3)，其濃度分別介於ND~249.05 ng/L、ND~1384.60 ng/L、ND~148.08 ng/L、ND~37.15 ng/L、ND~72.45 ng/L。
本研究結果化學分析與生物分析並沒有顯著的相關性，而根據化學分析與生物分析之結果進行相比較後發現，約有69%樣站其化學分析結果都高於生物分析結果，判斷可能是因為樣品經濃縮後產生的相關抑制效應或是樣品毒性導致有所差異；而在生物分析結果大於化學分析之樣站，由於水體存在天然雌激素如植物雌激素等，並沒有辦法忽略其效力。

Endocrine disrupting chemicals (EDCs) are commonly found in various industrial and household products. They enter the body by skin contact, inhalation, ingestion, or maternal exposure. Although they are found in very low concentrations, endocrine disorders such as developmental disorders, reproductive damage, obesity, cancer, and other diseases can be produced from different ways. Natural and synthetic estrogenic compounds are discharged into the environment through human and animal excreta. Their structures, receptor characteristics, and signal transduction characteristics are similar to those of estrogen, and they are therefore also known as xenoestrogens (XEs).
The Wuluo River, an important tributary of Taiwan’s second largest river, the Kuoping River, receives livestock wastewater and human sewage. Because of the high density of livestock along the river, it is important to understand the background estrogenic activity of animal husbandry excreta.
In this study, the water quality at twelve stations collected during different seasons in the Wuluo River Basin was investigated. In addition, T47D-Kbluc reporter gene assays and E-Screen assays were used to evaluate potential the estrogenicity of the collected samples. An HPLC-MS2 was used to analyze 22 compounds and their interrelationships were then determined.
Our results indicated that water quality was moderately to severely polluted during all four samplings. Bioassay results showed that there was high estrogenic activity in the waters of the Wuluo River and that it changes from wet to dry seasons and with extractions or not. Calculated estrogenic activity ranged from 11.66 and 190.06 ng/L in concentrated samples using T47D-Kbluc bioassays, which was higher than in non-concentrated raw water having estrogenic activity between 0.23 and 5.62 ng/L E2T47D-KblucEEQ. The concentration of E-Screen cell proliferation assays was between 6.63 ~ 84.50 ng/L E2E-ScreenEEQ. According to chemical analyses, compounds with higher detection rates greater than 85% were estrone (E1), bisphenol A (BPA), nonylphenol di-ethoxylate (NP2EO) and benzophenone-3 (BP-3). Their concentrations were ND ~ 249.05 ng/L, ND ~ 1384.60 ng/L, ND ~ 37.15 ng/L and ND ~ 72.45 ng/L, respectively.
There were no significant correlations between the chemical concentrations of target compounds and estrogenic activity using bioassays. Comparing the results obtained from chemical analyses and bioassays, we found that about 69% of the chemical analyses produced results that were higher than those of bioassays, which may be due to synergistic inhibitory effects or to sample toxicity caused by concentrating the samples. For those sites with higher estrogen potency than would be indicated from chemical analyses, we suggest that other estrogen-contributing compounds such as phytoestrogens should be further evaluated.

摘要 I
Abstract III
誌謝 V
縮寫表 VI
目錄 VII
圖目錄 X
表目錄 XII
1.前言 1
1.1 研究緣起 1
1.2研究目的 2
2.文獻回顧 3
2.1內分泌干擾物質(EDCs) 3
2.2目標化合物背景資料 11
2.2.1雌激素化合物 18
2.2.2防腐劑化合物 25
2.2.3防曬劑類化合物 30
2.2.4抗菌劑類化合物 33
2.2.5烷基酚類化合物 36
2.2.6驅蟲劑與興奮劑 40
2.3 研究地區背景 42
2.3.1氣候資料 42
2.3.2 水質資料 44
2.3.3 畜牧業現況 44
2.4 內分泌干擾物質化學分析 46
3.實驗假說 50
4.材料與方法 51
4.1 實驗材料 51
4.1.1 實驗藥品 51
4.1.2 實驗設備及器材 53
4.1.3 細胞株來源 54
4.2 樣品採集 55
4.2.1 採樣點環境 55
4.2.2 樣品採集 55
4.2.3 水質分析 55
4.3 化學分析 57
4.3.1 樣品前處理製備 57
4.3.2 參數設定 58
4.3.3 品保品管 60
4.4 生物分析 61
4.4.1 細胞培養 61
4.4.2 原水樣品製備 61
4.4.3 濃縮樣品製備 61
4.4.4 實驗流程 61
4.5 數據評估計算結果 65
4.5.1生物分析結果預估雌激素活性 65
4.5.2 CAFOs化學分析預估貢獻雌激素濃度 66
5.實驗結果 67
5.1 武洛溪水質資料 67
5.2 生物試驗 75
5.2.1 相關當量計算 75
5.2.2 T47D-Kbluc冷光報導基因法 75
5.2.3 E-Screen細胞增生法 76
5.2.2生物分析結果 78
5.3化學分析 83
5.4 預估的化學分析與生物分析評估綜合探討 90
5.4.1 數據評估前實際考量 90
5.4.2 預估的化學分析與T47D-Kbluc冷光報導基因法評估比較 90
5.4.3 預估的化學分析與E-Screen細胞增生法評估比較 91
5.5 化合物流佈與貢獻探討 95
5.6 預估CAFOs貢獻雌激素 98
5.7 雌激素類化合物季節性變化趨勢 99
6.結論與建議 103
7.參考文獻 112

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