臺灣博碩士論文加值系統

由於近年來工業科技的快速進步，許多內分泌干擾物質被廣泛應用於工業製程及日常用品，伴隨人類活動而大量釋放至環境中，其中包括芳香烴受體促進物質、類(抗)雄激素與類(抗)鹽皮質激素物質，當這些物質隨著廢水排放而進入污水處理廠，便可能在後續之加氯消毒流程產生氯化衍生物並排放至承受水體中，因此氯化衍生物之生成與其對水體生態可能之危害需要受到關注。
本研究以報導基因酵母菌生物試驗法分析臺灣五座污水處理廠各處理單元放流水中的芳香烴受體促進活性、類(抗)雄激素以及類(抗)鹽皮質激素活性，並利用液相層析串聯式質譜儀分析樣本中選定之EDCs與其氯化衍生物之濃度。生物試驗法之結果顯示各污水廠之樣本皆無檢測出類雄激素活性，而在所有水相以及懸浮固體相樣本中皆有檢出芳香烴受體促進活性與抗鹽皮質激素活性，抗雄激素活性則主要於水相樣本中檢出。
氯化衍生物之生物試驗結果顯示，對羥基苯甲酸甲酯之單氯衍生物具有顯著的芳香烴受體促進活性與抗雄激素活性，而其雙氯衍生物則僅誘發抗雄激素活性。對羥基苯甲酸丙酯氯化混合物能誘發較原物質更強之芳香烴受體促進活性與抗雄激素活性，三氯沙氯化混合物之芳香烴受體促進活性較三氯沙弱，而抗雄激素活性則略有增強。壬基酚氯化混合物依舊不具有芳香烴受體促進活性，且其誘發之抗雄激素活性有明顯增強之趨勢。
液相層析串聯式質譜儀之分析結果指出水相樣本中主要檢測出雙酚A (ND~923 ng/L)、雙氯雙酚A (ND~35.9 ng/L)、對羥基苯甲酸甲酯 (12.4~2422.7 ng/L)、單氯對羥基苯甲酸甲酯 (ND~26.9 ng/L)、三氯沙 (61.4~904.4 ng/L)以及壬基酚 (108.7~7715.8 ng/L)。此外，將水相樣本中經LC-MS/MS檢測出之各類EDCs濃度轉換為標準品當量濃度後與生物試驗之結果進行比較，可發現生物試驗所換算之標準品當量濃度遠高於儀器分析，可能之原因為環境樣本中含有之多種EDCs並未被選定為待測物質，因此其貢獻之活性未被計入，又或是樣本中複雜物質之組成產生協同作用而導致生物試驗所得活性偏高。
綜觀本研究之結果，此五座污水處理廠並無法完全去除內分泌干擾物質，甚至可能因後續加氯消毒流程而產生活性更強之氯化衍生物。未來針對環境中內分泌干擾物質氯化衍生物之影響與流佈值得更多研究關注。

There are various kinds of endocrine disrupting chemicals (EDCs) present in the effluent of wastewater treatment plants (WWTPs), such as chemicals showing disrupting activities for aryl hydrocarbon receptor (AhR), androgen receptor (AR), or mineralocorticoid receptor (MR). These EDCs may further react with chlorine to form chlorinated derivatives in disinfection process of WWTPs. In this study, samples from different treatment stages of WWTPs were collected and analyzed by recombinant yeast bioassays and LC-MS/MS to investigate the endocrine disrupting activities and concentrations of target EDCs and their chlorinated derivatives. Our results showed that AhR agonist activities, anti-androgenic activities, and anti-mineralocorticoid activities were detected frequently in the dissolved phase of WWTP influents and effluents. The chlorinated derivatives of selected EDCs still showed AhR agonist activities, AR antagonist activities, and MR antagonist activities in the bioassays, which were even stronger than their parent compounds. LC-MS/MS results showed that some EDCs, such as methylparaben, nonylphenol, and triclosan were often detected at ng/L to μg/L levels. In addition, chlorinated derivatives of methylparaben and bisphenol A were also often detected in WWTP samples. More attention should be paid to the formation of chlorinated EDCs and their potential risk to aquatic organisms.

摘要 I
Abstract III
致謝 VI
目錄 VII
表目錄 XI
圖目錄 XII
第一章 前言 1
1-1研究源起 1
1-2研究目的 2
第二章 文獻回顧 3
2-1 內分泌干擾物質 3
2-1-1 芳香烴受體促進物質 4
2-1-2 類(抗)雄激素物質 6
2-1-3 類(抗)鹽皮質激素物質 8
2-1-4 內分泌干擾物質之氯化衍生物 9
2-2 生物試驗方法 14
2-2-1 活體內生物試驗法 14
2-2-2 活體外生物試驗法 15
2-3儀器分析方法 16
2-3-1 層析系統 17
2-3-2 偵測系統 17
2-4檢測污水處理廠簡介 19
2-4-1臺南AP水資源回收中心 19
2-4-2臺南GT水資源回收中心 19
2-4-3臺南HW污水處理廠 19
2-4-4高雄NZ污水處理廠 20
2-4-5高雄FS污水處理廠 20
第三章 實驗方法與步驟 21
3-1 實驗流程 21
3-2 樣本採集 22
3-3 實驗材料與設備 24
3-3-1 藥品及試劑 24
3-3-2基因重組酵母菌試驗 26
3-3-3實驗用設備 28
3-4前處理步驟 29
3-4-1 水相樣本 29
3-4-2懸浮固體相樣本 29
3-4-3樣本稀釋序列 30
3-5基因重組酵母菌生物試驗法 31
3-5-1芳香烴受體促進活性試驗 31
3-5-2類(抗)雄激素活性試驗 33
3-5-3類(抗)鹽皮質激素活性試驗 34
3-5-4當量濃度換算與方法偵測 37
3-6層析儀器分析 38
3-6-1逆相高效液相層析儀 (RP-HPLC) 38
3-6-2液相層析串聯式質譜儀 (LC-MS/MS) 39
3-7 內分泌干擾物之氯化衍生物 43
3-7-1衍生物氯化合成與純化 43
第四章 結果與討論 45
4-1 芳香烴受體促進活性 45
4-1-1 AP水資源回收中心 45
4-1-2 GT水資源回收中心 46
4-1-3 HW污水處理廠 47
4-1-4 NZ污水處理廠 48
4-1-5 FS污水處理廠 49
4-1-6 水相樣本與懸浮固體相樣本中芳香烴受體促進活性之分布 50
4-1-7 各廠水處理單元之芳香烴受體促進活性綜合比較 51
4-1-8 各國污水廠水樣中芳香烴受體促進活性之比較 54
4-2 類(抗)雄激素活性 55
4-2-1 GT水資源回收中心 55
4-2-2 HW污水處理廠 56
4-2-3 NZ污水處理廠 57
4-2-4 FS污水處理廠 58
4-2-5 水相樣本與懸浮固體相樣本中抗雄激素活性之分布 59
4-2-6 各廠水處理單元之抗雄激素活性綜合比較 60
4-2-7 各國污水廠水樣中抗雄激素活性之比較 61
4-3 類(抗)鹽皮質激素活性 62
4-3-1 AP水資源回收中心 62
4-3-2 GT水資源回收中心 63
4-3-3 HW污水處理廠 64
4-3-4 NZ 污水處理廠 65
4-3-5 FS 污水處理廠 66
4-3-6 水相樣本與懸浮固體相樣本中抗鹽皮質激素活性分布 67
4-3-7 各廠水處理單元之抗鹽皮質激素活性綜合比較 68
4-4 生物試驗結果討論 71
4-4-1各污水廠樣本中類(抗)激素活性變化 71
4-5 EDCS 之氯化衍生物 75
4-5-1 氯化MeP之內分泌干擾活性 75
4-5-2 氯化PrP之內分泌干擾活性 77
4-5-3 氯化TCS之內分泌干擾活性 79
4-5-4 氯化NP之內分泌干擾活性 83
4-6 LC-MS/MS儀器分析結果討論 85
4-6-1污水處理廠中BPA濃度變化 85
4-6-2污水處理廠中氯化BPA濃度變化 87
4-6-3污水處理廠中MeP濃度變化 88
4-6-4污水處理廠中氯化MeP濃度變化 90
4-6-5污水處理廠中PrP濃度變化 91
4-6-6污水處理廠中TCS濃度變化 93
4-6-7污水處理廠中NP濃度變化 95
4-6-8 各國污水廠水樣中EDCs氯化衍生物濃度之比較 97
4-7 生物試驗與儀器分析結果比較 98
4-7-1水相樣本之生物試驗與儀器分析相關性 98
第五章 結論 102
5-1 結論 102
5-2 建議 104
參考文獻 105
附錄 116

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