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研究生： 江政哲
Cheng-Che Chiang
論文名稱： 以固相萃取法及氣相層析質譜儀對水環境中動情激素類有機污染物之分析與研究
Analysis of estrogens in environmental water samples using solid-phase extraction and gas chromatography–mass spectrometry
指導教授： 丁望賢
Wang-Hsien Ding
口試委員:
學位類別： 碩士
Master
系所名稱： 理學院 - 化學學系
Department of Chemistry
畢業學年度： 90
語文別： 中文
論文頁數： 67
中文關鍵詞： 水環境 、衍生化 、氣相層析質譜儀 、動情激素
外文關鍵詞： estrogen, gas chromatography mass spectrometry, derivatization
相關次數： 點閱：12  下載：0 
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    • 本研究藉著矽烷化衍生法將天然以及人工合成的動情激素類化合物衍生為低極性高揮發度的衍生物，同時搭配具有高靈敏度和良好分離效果的氣相層析質譜儀來對環境水樣作一分析。衍生化過程中藉著添加不同比例的催化劑(TMCS)至衍生試劑中(BSTFA)，來探討不同配方衍生試劑對最終衍生物的影響。實驗發現除了部分分析物無法完全衍生的主要因素是因為相鄰的OH官能基與乙炔基帶來的立體障礙影響，而隨著催化劑比例的增加也可以使具有2~3個OH官能基的分析物(例如：estriol , E3 ; 17α-ethynylestradiol , EE2)增加完全衍生的比例。在O-TMS衍生物的質譜圖中也發現，所有O-TMS的衍生物其強度最高的離子基峰皆為分子離子，因此為了求得在定性定量上最高的靈敏度，離子選擇主要皆以分子離子為主，另外部分碎裂較嚴重的衍生物，則再加上相對強度次高的特徵斷裂離子以加強積分訊號。
    萃取部分則是採用全新研發的Oasis® HLB固相萃取管柱，經由去離子水與真實水樣基質干擾的驗證下，回收率皆可以達到80﹪以上，而目前所抽查的四條台灣河流當中，由其分析結果得知目前台灣河流當中，天然與人工合成的動情激素類化合物含量皆低於本方法之偵測極限5 ppt。

    This work evaluated derivatization procedures for detecting both natural and synthetic estrogenic chemicals to obtain derivatives with high selectivity for using in gas chromatography-mass spectrometry (GC/MS) analysis of environmental samples. Different silylating agents, mainly trimethylsilylating agents (TMS), were compared, and the roles of various content of trimethylchlorosilane (TMCS, as a stimulator) were investigated. The difference in the abundances of the derivatives was caused by the sterical hindrance of multiple hydroxyl groups and ethynyl group in the structures of estrogenic chemicals. The use of TMCS produces an increase in the derivatization yield, especially for the compounds with multiple hydroxyl groups (i.e., estriol (E3) and 17a-ethynylestradiol (EE2)). Mass spectra of O-TMS-derivatives and tentative fragmentation profiles are also displayed. Molecular ions were the base peaks in all the derivatives, and were used as the quantitation ions to obtain maximum detection sensitivity and specificity. Sample enrichment was achieved by newly developed Oasisâ HLB solid-phase extraction cartridges. Recovery of the estrogenic chemicals in spiked various water samples ranged from 78% to 102% while RSD ranged from 1 to 15%.

    中文摘要 -------------------------------------------------------------- Ⅰ 英文摘要 -------------------------------------------------------------- Ⅱ 目錄 ------------------------------------------------------------------ Ⅲ 表目錄 ---------------------------------------------------------------- Ⅵ 圖目錄 ---------------------------------------------------------------- Ⅶ 第一章 前言 ---------------------------------------------------------- 1 1-1研究緣起 --------------------------------------------------------- 1 1-2研究目標 --------------------------------------------------------- 4 第二章 文獻回顧 ------------------------------------------------------ 6 2-1環境荷爾蒙簡介 --------------------------------------------------- 6 2-1-1環境荷爾蒙之作用機制 --------------------------------------- 7 2-1-2環境荷爾蒙物質對生態之影響 --------------------------------- 9 2-2動情激素類化合物 ------------------------------------------------- 12 2-2-1水環境中動情激素類化合物之來源 ----------------------------- 13 2-2-2動情激素類化合物之代謝變化 --------------------------------- 13 2-2-3國外相關研究 ----------------------------------------------- 15 2-3固相萃取法 ------------------------------------------------------- 16 2-4衍生化技術 ------------------------------------------------------- 18 2-4-1衍生化技術之條件 ------------------------------------------- 19 2-4-2矽烷化衍生法 ----------------------------------------------- 19 2-5氣相層析質譜儀 --------------------------------------------------- 21 2-5-1大體積直接進樣裝置 ----------------------------------------- 21 2-5-2離子阱質譜儀 ----------------------------------------------- 23 第三章 實驗步驟與樣品分析 -------------------------------------------- 26 3-1實驗藥品與儀器設備 ----------------------------------------------- 26 3-1-1實驗藥品 --------------------------------------------------- 26 3-1-2儀器設備 --------------------------------------------------- 28 3-2實驗步驟 --------------------------------------------------------- 29 3-2-1標準品的製備 ----------------------------------------------- 29 3-2-2標準品的測定 ----------------------------------------------- 31 3-2-3萃取步驟 --------------------------------------------------- 34 3-2-4衍生化步驟 ------------------------------------------------- 34 3-2-5真實水樣萃取及淨化 ----------------------------------------- 35 3-3樣品採集 --------------------------------------------------------- 35 第四章 結果與討論 ---------------------------------------------------- 36 4-1不同配方矽烷化試劑對衍生化產物之探討 ----------------------------- 36 4-1-1 BSTFA與MSTFA的TMS官能基團衍生反應--------------------------- 39 4-1-2立體障礙對MTBSTFA大體積TBDMS官能基團的影響 ------------------ 43 4-2時間對衍生化產率的探討 ------------------------------------------- 44 4-3動情激素類化合物之測定 ------------------------------------------- 45 4-4檢量線 ----------------------------------------------------------- 53 4-5測試動情激素類化合物於不同水環境中的萃取回收率 -------------------- 57 4-6台灣河川中動情激素類化合物的含量 --------------------------------- 59 第五章 結論與建議 ---------------------------------------------------- 61 5-1結論 ------------------------------------------------------------- 61 5-2建議 ------------------------------------------------------------- 62 參考文獻 -------------------------------------------------------------- 63 表目錄 表2-1. 常用的矽烷化衍生試劑 ------------------------------------------- 20 表3-1. DSI法注射埠溫度變化及分流-不分流之配合 ------------------------- 33 表4-1. 添加不同比例催化劑衍生試劑與其衍生產物的比例變化 --------------- 40 表4-2. 添加不同比例催化劑衍生試劑與其衍生產物的比例變化 --------------- 42 表4-3. 以電子撞擊游離法離子化的TMS衍生物特徵斷裂 ---------------------- 46 表4-4. 選擇積分之特徵離子斷裂 ----------------------------------------- 53 表4-5. TMS矽烷化標準品線性範圍與檢量線 -------------------------------- 56 表4-6. 不同基質中動情激素的回收率 ------------------------------------- 58 表4-7. 各個河水中動情激素類化合物之含量 ------------------------------- 60 圖目錄 圖1-1. 壬基苯酚(NP)及雌二醇(Estradiol)有相似的化學結構 ---------------- 1 圖1-2. 動情激素類有機污染物結構 --------------------------------------- 3 圖2-1. 荷爾蒙與荷爾蒙受體的「鎖和鑰匙」機制 --------------------------- 8 圖2-2. 美國安大略湖多氯聯苯的生物累積放大作用 ------------------------- 10 圖2-3. 雌性激素生物體內代謝過程 --------------------------------------- 14 圖2-4. 雄性激素生物體內代謝過程 --------------------------------------- 14 圖2-5. HLB吸附劑的分子結構 -------------------------------------------- 17 圖2-6. 固相萃取流程 --------------------------------------------------- 18 圖2-7. 大體積進樣裝置 ------------------------------------------------- 22 圖2-8. 離子阱質譜儀裝置簡圖 ------------------------------------------- 24 圖4-1. (a)E2的TMS單取代與雙取代衍生物 (b)E3的TMS雙取代與三取代衍生物 ---------------------------------- 37 圖4-2. (a)尚未衍生化(b)經TMS官能基團衍生化全離子層析圖 ---------------- 38 圖4-2. (c)經TBDMS官能基團衍生化全離子層析圖 --------------------------- 39 圖4-3. (a)E2 (b)E3 (c)EE2 (d)EA的TBDMS衍生物 -------------------------- 43 圖4-4. RF平均值對時間的關係圖 ----------------------------------------- 44 圖4-5. TMS矽烷化的標準品EI質譜圖(a)E1(b)4-OH E1 ----------------------- 48 圖4-6. TMS矽烷化的標準品EI質譜圖(a)E2(b)2-OH E2 ----------------------- 49 圖4-7. TMS矽烷化的標準品EI質譜圖(a)E3(b)TS ---------------------------- 50 圖4-8. TMS矽烷化的標準品EI質譜圖(a)EE2(b)MeEE2 ------------------------ 51 圖4-9. TMS矽烷化的標準品EI質譜圖(a)DES(b)EA --------------------------- 52 圖4-10. 標準品0.1 ppm的選擇離子層析圖(a) E1、4-OH E1、E2 -------------- 54 圖4-10. 標準品0.1 ppm的選擇離子層析圖 (b) 2-OH E2、E3、DES(c)EE2、MeEE2、TS --------------------------------- 55

    ◆王正雄：環境荷爾蒙-地球村二十一世紀之熱門課題，環境檢驗，第29期，第6-14頁，民國89年。
    ◆李美慧：常見環境荷爾蒙物質及其影響，環境荷爾蒙研討會論文集，台北生技中心，第4-13頁，民國89年。
    ◆何國榮、張耀仁：離子阱質譜儀，科儀新知，第14卷5期，第48-55頁，民國82年。
    ◆吳東傑、李芸玫、李靜瑤譯著，失竊的未來，先覺出版社，民國 89年。
    ◆陳文欽、楊琇塋、陳文卿：三丁錫對海洋生物的影響，第一屆環境荷爾蒙與持久性有機污染物研討會論文集，環境品質基金會，第87-91頁，民國89年。
    ◆黃忠智：氣相層析/離子阱串聯質譜儀，科儀新知，第17卷2期，第42-49頁，民國88年。
    ◆楊琇瑩：環境荷爾蒙物質對水生生物的影響，環境荷爾蒙研討會論文集，台北生技中心，第43-49頁，民國89年。
    ◆國科會精密儀器發展中心：低能量碰撞誘導解離之串聯質譜儀，質譜分析術專輯，民國81年。
    ◆Alda, M.J., Barcelo, D. (2000) Determination of Steroid Sex Hormones and Related Synthetic Compounds Considered as Endocrine Disrupters in Water by Liquid Chromatography-dioid Array Detection-mass Spectrometry, J. Chromatogr. A, 892, 391-406.
    ◆Belfroid, A.C., Horst, A.V., Vethaak, A.D., Schafer, A.J., Rijs, G.B.J., Wegener, J., Cofino, W.P. (1999) Sci. Total Environ., 225, 101-108.
    ◆Choi, M.H., Kim, K.R., Chung, B.C. (2000) Determaination of Estrogen and 17β-Estradiol in Human Hair by Gas Chromatography Mass Spectrometry, Analyst, 125, 711-714.
    ◆Ding, W.H., Tzing, S.H. (1998) Analysis of Nonylphenol Polyethoxylates and Their Degradation Products in River Water and Sewage Effluent by Gas Chromatography- Ion Trap (Tandem) Mass Spectrometry with Electron Impact and Chemical Ionization, J. Chromatogr. A, 824, 79-90.
    ◆Damasceno, L., Ventura, R., Ortuno, J., Segura, J. (2000) Derivatization Procedures for the Detection of β2-agonists by Gas Chromatographic/Mass Spectrometric Analysis, J. Mass Spectrom., 35, 1285-1294.
    ◆Harries, J.E., Sheahan, D.A., Jobling, S., Matthiessen, P., Neall, M., Sumpter, J.P., Taylor, T. and Zaman, N. (1997) Estrogenic Activity in Five United-Kingdom Rivers Detected by Measurement of Vitellogenesis in Caged Male Trout, Environ. Toxicol. Chem., 16, 534-542.
    ◆Jobling, S. and Sumpter, J.P. (1993) Detergent Components in Sewage Effluent are Weakly Estrogenic to Fish an-in-vitro Study Using Rainbow-Trout (Oncorhynchus-Mykiss) Hepatocytes, Aquat. Toxicol., 27, 361-372.
    ◆Kuch, H.M., Ballschmiter, K. (2001) Determination of Endocrine-Disrupting Phenolic Compounds and Estrogens in Surface and Drinking Water by HRGC－(NCI)－MS in the Picogram per Liter Range, Environ. Sci. Technol., 35, 3201-3206.
    ◆Kelly, C., (2000) Analysis of Steroids in Environmental Water Samples Using Solid-phase Extraction and Ion-trap Gas Chromatography Mass Spectrometry and Gas Chromatography Tandem Mass spectrometry, J. Chromatogr. A, 872, 309-314.
    ◆Lagana, A., Bacaloni, A., Fago, G., Marino, A. (2000) Trace Analysis of Estrogenic Chemicals in Sewage Effluent Using Liquid Chromatography Combined with Tandem Mass Spectrometry, Rapid Commun. Mass Spectrom., 14, 401-407.
    ◆Li, D., Park, J. (2001) Silyl Derivatization of Alkylphenols, Chlorophenols and Bisphenol A for Simultaneous GC/MS Determination, Anal. Chem., 73, 3089-3095.
    ◆Lee, H.B., Peart, T.E. (1998) Determination of 17β-Estradiol annd Its Metabolites in Sewage Effluent by Solid-phase Extraction and Gas Chromatography/Mass Spectrometry, J. AOAC International, 81, 1209-1216.
    ◆Nakamura, S., Sian, T.H., Daishima, S. (2001) Determination of Estrogens in river Water by Gas Chromatography－Negative-ion Chemical-ionization mass spectrometry, J. Chromatogr. A, 919, 275-282.
    ◆Oh, S.M., Choung, S.Y., Sheen, Y.Y., Chung, K.H. (2000) Quantitative Assessment of Estrogenic Activity in the Water Environment of Korea by the E-SCREEN Assay, Sci. Total Environ., 263, 161-169.
    ◆Ollers, S., Singer, H.P., Fassler, P., Muller, S.R. (2001) Simultaneous Quantification of Neutral and Acidic Pharmaceuticals and Pesticides at the low-ng/l level in Surface and waste and waste water, J. Chromatogr. A, 911, 225-234.
    ◆Pinnella, K.D., Cranmer, B.K., Tessari, J.D., Cosma, G.N., Veeramachaneni, D.N. (2001) Gas Chromatographic Detection of Catecholestrogens Following Isolation by Solid-phase Extraction, J. Chromatogr. B, 758, 145-152.
    ◆Purdom, C.E., Hardiman, P.A., Bye, V.J., Eno, N.C., Tyler, C.R., Sumpter, J.P. (1994) Chem. Ecol., 8, 275-285.
    ◆Snyder, S.A., Keith, T.L., Verbrugge, D.A., Snyder, E.M., Gross, T.S., Kannan, K., Giesy, J.P. (1999) Analytical Methods for Detection of Selected Estrogenic Compounds in Aqueous Mixtures, Environ. Sci. Technol., 33, 2814-2820.
    ◆Snyder, S.A., Villeneuve, D.L., Snyder, E.M., Giesy, J.P. (2001) Identification and Quantification of Estrogen Receptor Agonists in Wastewater Effluents, Environ. Sci. Technol., 35, 3620-3625.
    ◆Ternes, T.A., Stumpf, M., Mueller, J., Wilken, R.D., Servos, M. (1999) Behavior and Occurrence of Estrogens in Municipal Sewage Treatment Plants － I. Investigations in Germany, Canada and Brazil, Sci. Total Environ., 225, 81-90.
    ◆Waldock, M.J. and Thain, J.E. (1983) Shell Thickening in Crassostrea Gigs - Organotin Antifouling or Sediment Induced, Mar. Pollut. Bull., 14, 411-415.
    ◆Waters Oasis®, (2001) Sample Extraction : Environmental and Agrochemical Applications Notebook, Waters, Milford, MA, USA.
    ◆Xiao, X.Y., McCalley, D. (2000) Quantitative Analysis of Estrogens in Human Urine using Gas Chromatography/Negative Chemical Ionization Mass Spectrometry, Rapid Commun. Mass Spectrom., 14, 1991-2001.
    ◆Xiao, X.Y., McCalley, D.V., McEvoy, J. (2001) Analysis of Estrogens in River Water and Effluents Using Solid-phase Extraction and Gas Chromatography－Negative Chemical Ionization Mass Spectrometry of the Pentafluorobenzoyl Derivatives, J. Chromatogr. A, 923, 195-204.

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