臺灣博碩士論文加值系統

本研究旨在探討淨水場(板新、豐原、鳳山、福興等)處理前之原水與加氯消毒後之清水中有機物之致突變性與急毒性，針對溶解性有機碳、加氯消毒副產物(三鹵甲烷、鹵乙酸)與環境荷爾蒙(雙酚A、壬基酚)等水中有害物質，以化學定量分析其濃度，並利用生物毒性試驗法包括安姆測試法(Ames test)、姊妹子染色體交換測試法(Sister Chromatid Exchange, SCE)與螢光菌急毒性試驗法(Microtox test)檢測水質是否具有致突變性與急毒性，以建立其具實用性之分析程序，並整合化學與生物各項分析參數，探討自來水中有機物、環境荷爾蒙物質與加氯消毒副產物等之關連性。本研究結果如下:
1. 四個淨水場清水溶解性有機碳(DOC)在枯水期其測值為0.201~1.875mg/L，在豐水期其測值為0.296~0.647mg/L。
2. 四個淨水場清水三鹵甲烷(THMs)在枯水期其測值分別為氯仿<0.80~6.38μg/L、一溴二氯甲烷<0.80~2.75μg/L、二溴一氯甲烷<0.80~1.38μg/L、溴仿<0.80μg/L; 在豐水期其測值分別為氯仿<0.40~9.60μg/L、一溴二氯甲烷<0.40~4.95μg/L、二溴一氯甲烷<0.40~4.96μg/L、溴仿<0.40~2.05μg/L，皆符合飲用水水質標準80μg/L。
3. 鹵乙酸(HAAs)在枯水期其測值分別為二氯乙酸(DCAA) ND~4.71μg/L、三氯乙酸(TCAA) ND~27.02μg/L，其他則未檢出; 在豐水期其測值分別為二氯乙酸(DCAA) ND~6.50μg/L、三氯乙酸(TCAA) ND~5.22μg/L，其他則未檢出，因國內無其飲用水標準，但檢驗結果皆有符合美國環保署飲用水標準60μg/L。
4. 四個淨水場清水壬基酚(NP)在枯水期其測值為未檢出，在豐水期其測值為未檢出，雙酚A(BPA)在枯水期其測值為<0.005mg/L，在豐水期其測值為<0.005mg/L。
5. 壬基酚（Nonylphenol）、雙酚A（Bisphenol-A）原體不誘發TA98、TA100之致突變；三氯乙酸（CHCl3COOH），溴乙酸（CH2BrCOOH）、二氯乙酸（CHCl2COOH）原體可誘發TA98之單一劑量之致突變性，但對TA100均無致突變性。氯乙酸（CH2ClCOOH）原體對TA98、TA100在特定劑量下可產生突變性，三氯甲烷（CHCl3）、二溴一氯甲烷（CHClBr2）原體對TA98及TA100均不產生致突變性。
6. 壬基酚（Nonylphenol）、雙酚A（Bisphenol-A）原體在濃度<500mg/L時對倉鼠卵細胞之姊妹子染色體無致變異性產生。
7. 2009年枯水期及豐水期之台灣四個自來水淨水場之原、清水利用沙門菌TA98及TA100均不產任何致突變性。
8. 2009年枯水期及豐水期之台灣四個自來水埸之原、清水利用倉鼠細胞之姊妹子染色體交換測試均不產生染色體變異性。
9. 利用螢光偵測儀（Microtox）測試顯示雙酚A及溴化物類原體的毒性較三鹵甲烷以及壬基酚強。
10. 利用螢光偵測儀（Microtox）測試台灣2009年枯水期及豐水期之四個自來水淨水場之原、清水，顯示各淨水場之水質屬於安全範圍內。
11. 本實驗之三種自來水污染物快速檢測法中，以Microtox最為迅速、簡便，其次是安姆氏試驗 ，姊妹子染色體交換法稍微複雜，本實驗建議，自來水生物檢測可利用Microtox方法搭配安姆氏試驗方法，可求得快速、正確之生物檢測結果。

This paper aims to investigate the mutation and acute toxicity of raw water and purified water of four water purification station including Ben-Sin, Fong-Yuan, Fong-Shan and Fu-Sing. Chemical analysis of water contained basic parameters, dissolved organic carbon, disinfection by-products of purified water such as trihalomethanes (THMs) and haloacetic acids (HAAs) and environmental hormones such as Bisphenol A (BPA) and Nonylphenol (NP). On the other hand, biological examination such as Ames test, Sister chromatid exchange (SCE) and Microtox test were conducted to test the potential mutation effects due to the exposure of chemical contaminants potentially exisisting in water. Correlations of chemical contaminants analysis and the biological tests were examined to provide the baseline information for the operation and management of water supply plant. Results of this study were described as follows:
1. DOC was measured to be 0.201~1.875mg/L in the rain scarce season while it was analyzed to be 0.296~0.647 mg/L in the rain rich season respectively.
2. CHCl3, CHCl2Br, CHClBr2 and CHBr3 were &lt;0.80~6.38, &lt;0.80~2.75, &lt;0.80~1.38 and &lt;0.80 μg/L in the rain scarce season while they were &lt;0.40~9.60, &lt;0.40~4.95, &lt;0.40~4.96 and &lt;0.40~2.05 μg/L in the rain rich season respectively. Those measured values all meet the Taiwan regulatory standard of 80μg/L.
3. DCAA and TCAA were ND~4.71 μg/L and ND~27.02 μg/L respectively in the rain scarce season while those were ND~6.50 μg/L and ND~5.22 μg/L in the rain rich season respectively. Other compounds of HAAs were not measured (below MDL) in the study. Measured values meet the US EPA HAAs regulatory standard of 60μg/L although the HAAs standard were not promulgated at present in Taiwan.
4. NP were not found in rain scarce and rain rich season. BPA had lower values less than &lt;0.005 mg/L in rain scarce and rain rich season.
5. NP and BPA did not induce the mutation of TA98 and TA100; CHCl3COOH, CH2BrCOOH and CHCl2COOH compounds could induce the TA98 mutation while it did not induce TA100 mutation. CHCl3 and CHClBr2 did not induce mutation for both TA98 and TA100.
6. NP and BPA less than 500 mg/L did not cause the mutation of Chinese Hamster Ovary (CHO).
7. Raw and purified water did not induce TA98 and TA100 mutation in dry and rain rich season in 2009.
8. Raw and purified water did not induce CHO chromosome mutation in the dry and rain rich season in 2009.
9. Microtox test results showed that the toxicity of BPA and Bromide compounds was higher than those of THMs and NP ones.
10. Microtox test of raw and purified water showed that water quality was in the safe range in the rain scarce and rain rich season in 2009.
11. Microtox test was found to be simple and convenient among the three analysis methods. Ames test was the second and SCE was thought to be more complicated and time consuming. It is recommended that the use of Microtox test combined with Ames test could obtain the quick and precise results.

目錄
表目錄 XIV
圖目錄 XVII
第一章 研究主旨 1
第二章 研究緣起 2
2.1研究緣起 2
2.2 背景分析 3
2.3 水中毒性物質 8
第三章 文獻回顧 9
3.1 消毒副產物（Disinfection by products, DBPs） 9
3.1.1 影響DBPs生成之因素 9
3.1.1.1 前驅物質(Precursors) 10
3.1.1.2 有機物的含量 11
3.1.1.3 季節性變化 12
3.1.1.4 加氯量 12
3.1.1.5 pH值 12
3.1.2 DBPs定義 13
3.1.3 消毒副產物種類 14
3.1.4 三鹵甲烷(Trihalomethanes, THMs) 17
3.1.5 鹵乙酸(Haloacetic acids, HAAs) 23
3.1.6 消毒副產物的危害性 26
3.2 環境荷爾蒙((Environmental hormones, EHs) 27
3.2.1 雙酚A(Bisphenol A, BPA) 29
3.2.2 壬基酚(Nonylphenol) 31
3.3 致突變性與急毒性 34
3.3.1 安姆氏試驗基因毒性試驗基本原理 34
3.3.2 姊妹子染色體交換測試法原理 35
3.3.3 Microtox螢光菌急毒性試驗 35
3.3.3.1 Microtox 測試原理 35
3.3.3.2 Microtox之優點 36
3.4總結 37
第四章 研究方法及步驟 39
4.1 研究流程規劃 39
4.2 水樣採集分析 40
4.2.1 採樣地點與頻率 40
4.2.2 採樣設備與儲存 41
4.2.3 採樣分析品保目標 43
4.2.3.1 採樣程序 44
4.3 基本分析 52
4.3.1 pH/ORP 52
4.3.2 溶氧(DO) 52
4.3.3 導電度(EC) 52
4.4 化學分析－消毒副產物 53
4.4.1 三鹵甲烷分析(NIEA W781.50A) 53
4.4.2 鹵乙酸分析(NIEA W538.50C) 57
4.5 環境荷爾蒙分析 63
4.6生物檢測方法－安姆氏試驗 67
4.6.1 試驗材料 67
4.6.1.1 培養基 67
4.6.1.2 軟性瓊脂（Soft agar） 67
4.6.1.3 0.5 mM Histidine/biotin 溶液 67
4.6.1.4 0.1 M Histidine 組胺酸溶液 67
4.6.1.5 Ampicillin 溶液 67
4.6.1.6 Crystal violet 溶液 68
4.6.1.7 青黴素培養基（Ampicillin plate） 68
4.6.1.8 S9混合液製備 68
4.6.1.9 菌種培養 70
4.6.2 試驗設備 71
4.6.3 試驗方法 71
4.6.3.1 供試藥劑之處理 71
4.6.3.2 斑點測試 72
4.6.3.3 平板混合測試 73
4.6.4 結果分析 73
4.6.4.1 基因變異反應參考對照 73
4.6.4.2 正反應判定 74
4.6.4.3 負反應判定 74
4.6.5 試驗品保 75
4.6.5.1 整體結果 75
4.6.5.2 正反應組 75
4.6.5.3 負反應組 75
4.7 SCE測試方法 75
4.7.1 儀器設備及材料 75
4.7.2 試驗藥劑 77
4.7.3 試驗方法 78
4.7.3.1 藥劑配製 78
4.7.3.2 細胞計算法 79
4.7.3.3 細胞解凍 79
4.7.3.4 細胞繼代培養備製 79
4.7.3.5 繼代培養細胞保存 80
4.7.3.6 S9製作方法 80
4.7.4 試驗流程 81
4.7.4.1 試驗前3天將中國倉鼠細胞於T75培養瓶中解凍 81
4.7.4.2 經過3天培養後，將細胞分植培養於T25培養瓶中 81
4.7.4.3 加藥處理 82
4.7.5 加藥處理之優缺點 84
4.7.6 姊妹子染色體交換變異性反應試驗對照 84
4.7.6.1 空白對照 84
4.7.6.2 負反應對照 84
4.7.6.3 正反應對照 84
4.7.7 結果分析 85
4.7.7.1 對照之分析 85
4.7.7.2 劑量組與空白組之間之分析 85
4.7.8 試驗品保 85
4.8螢光菌急毒性試驗－Microtox 86
4.8.1 儀器設備 86
4.8.2 試驗材料 86
4.8.3 試驗方法 87
4.8.4 試驗流程 88
4.8.5 結果分析 90
4.8.6 Microtox試驗之優點 91
4.8.7 Microtox試驗品保 91
4.8.8 Microtox應用範圍 91
第五章 結果與討論 93
5.1 三鹵甲烷 93
5.2 鹵乙酸 94
5.3壬基酚與雙酚A 95
5.4 pH、ORP、DOC、餘氯、重金屬 97
5.5生物檢測 99
5.6給水處理方法與風險評估 118
5.7化學與生物分析相關性 120
5.8 毒理資料庫 121
第六章 結論與建議 126
6.1 結論 126
6.2建議 127
第七章 參考文獻 128

表目錄
表3-1 影響DBPs生成因子 10
表3-2 天然有機物與消毒副產物之相關性 14
表3-3 消毒副產物之分類 15
表3-4 三鹵甲烷致癌性風險評估 16
表3-5 各地區對DBPs之相關規範 17
表3-6 消毒副產物之三鹵甲烷致突變性 19
表3-6 消毒副產物之三鹵甲烷致突變性(續) 20
表3-7 三鹵甲烷對齧齒目動物之致癌性 21
表3-7 三鹵甲烷對齧齒目動物之致癌性(續) 22
表3-8 鹵乙酸類之物種、中英文名稱及化學式 23
表3-9消毒副產物鹵乙酸之致突變性 26
表3-10 雙酚A基本物化特性 30
表3-11 壬基酚基本物化特性 32
表3-12 細菌性生物毒性試驗的特色 37
表4-1 採樣所需設備藥劑 42
表4-2 樣品儲存方法 42
表4-3化學檢測數據品保目標 43
表4-4生物檢測數據品保目標 44
表4-5水質採樣設備使用及行前檢查表 46
表4-6樣品標籤 47
表4-7現勘紀錄表 48
表4-8樣品封條 49
表4-9樣品保存運送、接收紀錄表 50
表4-10樣品保存運送、接收紀錄表 51
表4-11 分析三鹵甲烷所需之藥劑與設備 54
表4-12 捕捉濃縮之建議條件 55
表4-13氣相層析儀與電子捕捉偵測器條件 55
表4-14 鹵乙酸各化合物之偵測極限 57
表4-15分析鹵乙酸所需之藥劑與設備 58
表4-16分析鹵乙酸所需之藥劑與設備(續) 59
表4-17 GC/ECD建議之操作條件 60
表4-18環境荷爾蒙分析所需設備與藥劑 64
表4-19環境荷爾蒙分析所需設備與藥劑(續) 65
表4-20 液相層析質譜儀參數設定： 66
表4-21每公升培養基含成分 68
表4-22混合液每mL所含成分 69
表4-23 加藥處理之優缺點 84
表5-1 調查場址枯水期原水與清水之三鹵甲烷分析濃度 93
表5-2 調查場址豐水期原水與清水之三鹵甲烷分析濃度 94
表5-3 調查場址枯水期原水與清水之鹵乙酸分析濃度 95
表5-4 調查場址豐水期原水與清水之鹵乙酸分析濃度 95
表5-5 調查場址枯水期與豐水期原水與清水之壬基酚及雙酚A濃度 96
表5-6西班牙原水雙酚A經各淨水單元處理後之濃度變化 96
表5-7 調查場址枯水期原水與清水之pH值 97
表5-8 調查場址豐水期原水與清水之pH值 97
表5-9 調查場址枯水期原水與清水之DOC濃度 98
表5-10 調查場址豐水期原水與清水之DOC濃度 98
表5-11 調查場址枯水期原水與清水之重金屬濃度 98
表5-12 調查場址豐水期原水與清水之重金屬濃度 98
表5-13 壬基酚(Nonyl phenol對沙門氏菌TA98誘發之變異菌落數 103
表5-14壬基酚(Nonyl phenol)對沙門氏菌TA100誘發之變異菌落數 103
表5-15 雙酚A(Bisphenol)對沙門氏菌TA98誘發之變異菌落數 104
表5-16 雙酚A(Bisphenol)對沙門氏菌TA100誘發之變異菌落數 104
表5-17 三氯乙酸(CHCl3COOH,TCAA)對沙門氏菌TA98誘發之變異菌落數 105
表5-18三氯乙酸(CHCl3COOH,TCAA)對沙門氏菌TA100誘發之變異菌落數 105
表5-19 溴乙酸(CH2BrCOOH,MBAA)對沙門氏菌TA98誘發之變異菌落數 106
表5-20 溴乙酸(CH2BrCOOH,MBAA)對沙門氏菌TA100誘發之變異菌落數 106
表5-21氯乙酸(CH2ClCOOH,MCAA)對沙門氏菌TA98誘發之變異菌落數 107
表5-22氯乙酸(CH2ClCOOH,MCAA)對沙門氏菌TA100誘發之變異菌落數 107
表5-23二氯乙酸(CHCl2COOH,DCAA)對沙門氏菌TA98誘發之變異菌落數 108
表5-24 二氯乙酸(CHCl2COOH,DCAA)對沙門氏菌TA100誘發之變異菌落數 108
表5-25 三氯甲烷(CHCl3)對沙門氏菌TA98誘發之變異菌落數 109
表5-26 三氯甲烷(CHCl3)對沙門氏菌TA100誘發之變異菌落數 109
表5-27 二溴一氯甲烷對沙門氏菌TA98誘發之變異菌落數 110
表5-28 二溴一氯甲烷對沙門氏菌TA100誘發之變異菌落數 110
表5-29 壬基酚對倉鼠卵巢細胞姊妹子染色體交換之變異數 111
表5-30 雙酚A對倉鼠卵巢細胞姊妹子染色體交換之變異數 111
表5-31 2009年枯水期台灣四個自來水淨水場之原水及清水對沙門氏菌TA98及TA100之變異菌落數 112
表5-32 2009年豐水期台灣四個自來水淨水場之原清水對沙門氏TA98 及TA100之變異菌落數. 112
表5-33 2009年枯水期台灣四個自來水淨水場之原清水對倉鼠卵細胞之姊妹子染色體交換之交換數 113
表5-34 2009年豐水期台灣四個自來水淨水場之原清水對倉鼠卵細胞之姊妹子染色體交換之交換數 113
表5-35利用螢光偵測法(Microtox)於2009年偵測台灣四個自來水淨水場枯水期之各水場原、清水樣本之EC50及EC20值 114
表5-36利用螢光偵測法(Microtox)於2009年偵測台灣四個自來水淨水場豐水期之各水場原、清水樣本之EC50及EC20值 114
表5-37大陸水樣經各淨水單元處理後之安姆氏試驗結果 115
表5-38經曝露處理BPA與NP 3小時之SCE值 116
表5-39義大利給淨水場3取水站D之安姆氏試驗 測試結果 117
表5-40義大利四淨水場中取水站A, B, C, D之Microtox test 測試結果 117
表5-41三鹵甲烷之安姆氏試驗 測試結果 118
表5-42大陸與加拿大給淨水場飲用水之致癌風險潛勢 119
表5-43台灣北中南地區給淨水場飲用水之致癌風險潛勢 119
表5-44台灣各地區給淨水場飲用水之致癌風險潛勢 120
表5-45利用螢光偵測法(Microtox)測試壬基酚、雙酚A及三鹵甲烷及溴乙酸類原體藥物之EC50及EC20值 121
表5-46利用螢光偵測法(Microtox)測試重金屬標準品之EC50及EC20值 123


圖目錄
圖2-1 板新淨水場淨水處理流程 3
圖2-2豐原淨水場淨水處理流程 4
圖2-3鳳山淨水場淨水處理流程 5
圖2-4福興淨水場淨水處理流程 5
圖3-1 三鹵甲烷結構式 18
圖3-2 US EPA規範之五種鹵乙酸結構式 24
圖3-3 雙酚A之化學結構式 30
圖3-4 環氧樹脂之化學結構式 31
圖3-5 壬基酚類持久性有機污染物結構 32
圖4-1 研究流程規劃圖 40
圖4-2 採樣流程圖 41
圖4-3採樣作業程序流程圖 45
圖4-4 基本分析流程圖 52
圖4-5 HAAs(GC/ECD)水樣萃取流程圖 61
圖4-6 酸性甲醇進行樣品甲基化反應步驟流程圖 62
圖4-7安姆氏（Ames test）基因變異試驗試驗設備 71
圖4-8斑點測試流程圖 72
圖4-9平板混合測試流程圖 73
圖4-10姊妹子染色體交換(SCE)試驗設備 77
圖4-11毒性物質分析儀 86
圖4-12螢光生物毒性(Microtox)檢測設備 87
圖4-13 Microtox試驗流程圖 89
圖5-1 二氯一溴甲烷原體藥物之螢光菌生物毒理反應曲線 122
圖5-2 三氯乙酸初始濃度40 mg/L暴露之螢光菌生物毒理反應曲線 122
圖5-3 Cr標準品之螢光菌生物毒理反應曲線 123
圖5-4 Ni標準品之螢光菌生物毒理反應曲線 124
圖5-5 Fe標準品之螢光菌生物毒理反應曲線 124
圖5-6 Zn標準品之螢光菌生物毒理反應曲線 125

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