Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/14589
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dc.contributor李茂榮zh_TW
dc.contributorMaw-Rong Leeen_US
dc.contributor.author尤婉柔zh_TW
dc.contributor.authorYu, Wan-Jouen_US
dc.contributor.other化學系所zh_TW
dc.date2013en_US
dc.date.accessioned2014-06-06T06:52:47Z-
dc.date.available2014-06-06T06:52:47Z-
dc.identifierU0005-3007201317504600en_US
dc.identifier.citation1. Qin, L. Q.; Wang, P. Y.; Kaneko, T.; Hoshi, K.; Sato, A., Estrogen: one of the risk factors in milk for prostate cancer. Med Hypotheses 2004, 62, 133-142. 2. 蔣春燕、張曉菊、應月青合著。「乳及乳制品中性激素殘留的安全現狀及檢測方法」。中國乳業 (2011),頁 59-61。 3. Farke, C.; Rattenberger, E.; Roiger, S. U.; Meyer, H. H., Bovine colostrum: determination of naturally occurring steroid hormones by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). J. Agric. Food Chem. 2011, 59, 1423-7. 4. Hanselman, T. A.; Graetz, D. A.; Wilkie, A. C., Manure-borne estrogens as potential environmental contaminants: A review. Environ Sci Technol. 2003, 37, 5471-5478. 5. Isobe, T.; Shiraishi, H.; Yasuda, M.; Shinoda, A.; Suzuki, H.; Morita, M., Determination of estrogens and their conjugates in water using solid-phase extraction followed by liquid chromatography-tandem mass spectrometry. J. Chromatogr. A 2003, 984, 195-202. 6. Tso, J.; Aga, D. S., Wrong-way-round ionization of sulfonamides and tetracyclines enables simultaneous analysis with free and conjugated estrogens by liquid chromatography tandem mass spectrometry. Anal. Chem. 2011, 83, 269-277. 7. Okoh, V.; Deoraj, A.; Roy, D., Estrogen-induced reactive oxygen species-mediated signalings contribute to breast cancer. Biochim. Biophys. Acta 2011, 1815, 115-33. 8. Xu, X.; Veenstra, T. D.; Fox, S. D.; Roman, J. M.; Issaq, H. J.; Falk, R.; Saavedra, J. E.; Keefer, L. K.; Ziegler, R. G., Measuring fifteen endogenous estrogens simultaneously in human urine by high -performance liquid chromatography-mass spectrometry. Anal. Chem. 2005, 77, 6646-6654. 9. 沈珺、徐瑩合著。「淺論牛乳中的雌激素」。黑龍江科技信息(2011),頁 184。 10. 姚曉芬、朱婧、楊月欣合著。「食品及母乳中雌性激素含量分析」。衛生研究( 2011),頁799-801。 11. 朱美鳳。「乳牛 TMR 飼料配方線上計算系統」。國立中興大學動物科學系,碩士論文,民96年。 12. Courant, F.; Antignac, J. P.; Laille, J.; Monteau, F.; Andre, F.; Le Bizec, B., Exposure assessment of prepubertal children to steroid endocrine disruptors. 2. Determination of steroid hormones in milk, egg, and meat samples. J. Agric. Food Chem. 2008, 56, 3176-3184. 13. McLachlan, J. A.; Simpson, E.; Martin, M., Endocrine disrupters and female reproductive health. Best Pract Res Cl En 2006, 20, 63-75. 14. Parodi, P. W., Impact of cows’ milk estrogen on cancer risk. Int. Dairy J. 2012, 22, 3-14. 15. Socas-Rodriguez, B.; Asensio-Ramos, M.; Hernandez-Borges, J.; Herrera-Herrera, A. V.; Rodriguez-Delgado, M. A., Chromatographic analysis of natural and synthetic estrogens in milk and dairy products. TrAC, Trends Anal. Chem. 2013, 44, 58-77. 16. Malekinejad, H.; Scherpenisse, P.; Bergwerff, A. A., Naturally occurring estrogens in processed milk and in raw milk (from gestated cows). J. Agric. Food Chem. 2006, 54, 9785-9791. 17. Shao, B.; Zhao, R.; Meng, J.; Xue, Y.; Wu, G.; Hu, J.; Tu, X., Simultaneous determination of residual hormonal chemicals in meat, kidney, liver tissues and milk by liquid chromatography–tandem mass spectrometry. Anal. Chim. Acta 2005, 548, 41-50. 18. Tso, J.; Aga, D. S., A systematic investigation to optimize simultaneous extraction and liquid chromatography tandem mass spectrometry analysis of estrogens and their conjugated metabolites in milk. J. Chromatogr. A 2010, 1217, 4784-95. 19. Wang, H.-X.; Zhou, Y.; Jiang, Q.-W., Simultaneous analysis of nine estrogens in milk powder with ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. Chin. J. Anal. Chem. 2011, 39, 1323-1328. 20. Yan, W.; Li, Y.; Zhao, L.; Lin, J. M., Determination of estrogens and bisphenol A in bovine milk by automated on-line C30 solid-phase extraction coupled with high-performance liquid chromatography-mass spectrometry. J. Chromatogr. A 2009, 1216, 7539-45. 21. Yang, Y.; Shao, B.; Zhang, J.; Wu, Y.; Duan, H., Determination of the residues of 50 anabolic hormones in muscle, milk and liver by very-high-pressure liquid chromatography-electrospray ionization tandem mass spectrometry. J. Chromatogr. B 2009, 877, 489-96. 22. Gabet, V.; Miege, C.; Bados, P.; Coquery, M., Analysis of estrogens in environmental matrices. TrAC, Trends Anal. Chem. 2007, 26, 1113-1131. 23. LaFleur, A. D.; Schug, K. A., A review of separation methods for the determination of estrogens and plastics-derived estrogen mimics from aqueous systems. Anal. Chim. Acta 2011, 696, 6-26. 24. Azzouz, A.; Jurado-Sanchez, B.; Souhail, B.; Ballesteros, E., Simultaneous determination of 20 pharmacologically active substances in cow''s milk, goat''s milk, and human breast milk by gas chromatography-mass spectrometry. J. Agric. Food Chem 2011, 59, 5125-32. 25. Markopoulou, C.K.; Koundourellis, J.E., Development of a validated liquid chromatography method for the simultaneous determination of ethinyl estradiol, cyproterone acetate, and norgestrel in breast milk following solid-phase extraction J. Liq. Chromatogr. Relat. Technol 2006, 29 , 685 26. Harwood, D. T.; Handelsman, D. J., Development and validation of a sensitive liquid chromatography-tandem mass spectrometry assay to simultaneously measure androgens and estrogens in serum without derivatization. Clin. Chim. Acta 2009, 409, 78-84. 27. Kushnir, M. M.; Rockwood, A. L.; Bergquist, J.; Varshavsky, M.; Roberts, W. L.; Yue, B.; Bunker, A. M.; Meikle, A. W., High-sensitivity tandem mass spectrometry assay for serum estrone and estradiol. Am J Clin Pathol. 2008, 129, 530-9. 28. Tai, S. S. C.; Welsh, M. J., Development and evaluation of a reference measurement procedure for the determination of estradiol-17 beta in human serum using isotope-dilution liquid chromatography-tandem mass spectrometry. Anal. Chem. 2005, 77, 6359-6363. 29. Lin, Y. H.; Chen, C. Y.; Wang, G. S., Analysis of steroid estrogens in water using liquid chromatography/tandem mass spectrometry with chemical derivatizations. Rapid Commun. Mass Spectrom. 2007, 21, 1973-83. 30. Nelson, R. E.; Grebe, S. K.; DJ, O. K.; Singh, R. J., Liquid chromatography-tandem mass spectrometry assay for simultaneous measurement of estradiol and estrone in human plasma. Clin. Chem. 2004, 50, 373-84. 31. Santa, T., ; Al-Dirbashi, O. Y ,; Fukushima,T., Derivatization reagents in liquid chromatography/electrospray ionization tandem mass spectrometry for biomedical analysis. Drug Discov Ther 2007, 2, 108-118. 32. Li, W.; Li, Y. H.; Li, A. C.; Zhou, S.; Naidong, W., Simultaneous determination of norethindrone and ethinyl estradiol in human plasma by high performance liquid chromatography with tandem mass spectrometry--experiences on developing a highly selective method using derivatization reagent for enhancing sensitivity. J. Chromatogr. B 2005, 825, 223-32. 33. 李茂榮。「低能量碰撞誘導解離之串聯質譜儀」。質譜分析技術專輯,民國91年,行政院國家科學委員會精密儀器發展中心編印,頁87-111。 34. Dass, C. Fundamentals of contemporary mass spectrometry; John Wiley & Sons, 2007, p.53. 35. 林秋束。「液相層析質譜術於尿液中鎮靜安眠類及濫用藥物之偵測」。國立中興大學化學研究所,碩士論文,民97年。 36. FinniganTM H-ESITM Probe User Guide, 2009, p.2. 37. FinniganTM Ion MaxTM and Ion Max-STM API Source Hardware Manual, 2008, p.58-59. 38. 李茂榮。「串聯質譜法(MS/MS)之原理與應用」。科儀新知58期(1990),頁8-19。 39. Louch, D.; Motlagh, S.; Pawliszyn, J., Dynamics of organic-compound extraction from water using liquid-coated fused-silica fibers. Anal. Chem. 1992, 64, 1187-1199. 40. 葉曜嘉。「固相微萃取法於微量氯酚之萃取偵測」。國立中興大學化學研究所,碩士論文,民85年。 41. 陸冠輝。「固相微萃取結合液相層析質譜儀於磺胺類劑之檢測」。國立中興大學化學研究所,碩士論文,民92年。 42. Kataoka, H.; Lord, H. L.; Pawliszyn, J., Applications of solid-phase microextraction in food analysis. J. Chromatogr. A 2000, 880, 35-62. 43. 李茂榮、陳崇宇、李祖光合著。「固相微萃取技術於微量分析之應用」。化學,第63卷,第3期(2005),頁329-342。 44. SUPELCO, SUPELCO Chromatography product, (1998) 325. 45. Musteata, M. L.; Musteata, F. M.; Pawliszyn, J., Biocompatible solid-phase microextraction coatings based on polyacrylonitrile and solid-phase extraction phases. Anal. Chem. 2007, 79, 6903-6911. 46. Vuckovic, D.; Pawliszyn, J., Systematic evaluation of solid-phase microextraction coatings for untargeted metabolomic profiling of biological fluids by liquid chromatography-mass spectrometry. Anal. Chem. 2011, 83, 1944-54. 47. Zhang, X.; Oakes, K. D.; Luong, D.; Metcalfe, C. D.; Servos, M. R., Solid-phase microextraction coupled to LC-ESI-MS/MS: evaluation and correction for matrix-induced ionization suppression/enhancement for pharmaceutical analysis in biological and environmental samples. Anal. Chem. 2011, 83, 6532-8. 48. 潘加亮、胡玉玲、李攻科合著。「生物相容性分離介質在樣品前處理中的應用」。分析化學 (2009),頁1539-1546。 49. Vuckovic, D.; Shirey, R.; Chen, Y.; Sidisky, L.; Aurand, C.; Stenerson, K.; Pawliszyn, J., In vitro evaluation of new biocompatible coatings for solid-phase microextraction: implications for drug analysis and in vivo sampling applications. Anal. Chim. Acta 2009, 638, 175-85. 50. Souverain, S.; Rudaz, S.; Veuthey, J. L., Restricted access materials and large particle supports for on-line sample preparation: an attractive approach for biological fluids analysis. J. Chromatogr. B 2004, 801, 141-156. 51. Lord, H. L.; Grant, R. P.; Walles, M.; Incledon, B.; Fahie, B.; Pawliszyn, J. B., Development and evaluation of a solid-phase microextraction probe for in vivo pharmacokinetic studies. Anal. Chem. 2003, 75, 5103-5115. 52. Olszowy, P.; Szultka, M.; Ligor, T.; Nowaczyk, J.; Buszewski, B., Fibers with polypyrrole and polythiophene phases for isolation and determination of adrenolytic drugs from human plasma by SPME-HPLC. J. Chromatogr. B 2010, 878, 2226-34.en_US
dc.identifier.urihttp://hdl.handle.net/11455/14589-
dc.description.abstract本研究利用生物相容性之固相微萃取技術(biocompatible solid-phase microexaction)結合衍生化之前處理方式,藉由液相層析串聯質譜術(LC-MS/MS)之選擇反應偵測模式進行牛奶中五種雌激素檢測。所分析化合物包括雌酮、17α-雌二醇、雌二醇、雌三醇和17α-乙炔基雌脂二醇等五種雌激素。研究中探討萃取溶液 pH 值、鹽類添加量、萃取時間、脫附溶劑、脫附時間等固相微萃取參數最佳化。 實驗結果顯示,固相微萃取 C18 纖維在水/甲醇(1/1,v/v)活化 10 分鐘後,牛奶不需經過蛋白質沉澱步驟,直接將纖維浸在牛奶中萃取 30 分鐘,於 1.5 毫升乙腈脫附 10 分鐘後,於衍生化試劑丹磺醯氯衍生化後分析,可得到良好靈敏度及選擇性。五種分析物線性範圍為 0.1 至 50 ng/mL,線性相關係數皆在 0.9956 以上,定量極限 6.1 至 14.0 pg/mL。所開發分析方法之同日內精確度與異日間精確度其相對標準偏差(relative standard deviation, RSD)分別介於 1.5 至 15.7% 與 3.9 至 18.2%之間;回收率則介於 90.1% 至 113.9% 之間。此分析方法減少對複雜基質前處理步驟、萃取效率高且對分析物具有選擇性。應用於市售牛奶中微量雌激素之檢測,實驗結果顯示並未檢測出含有雌激素之真實樣品。zh_TW
dc.description.abstractThe study proposes a novel method to determine trace estrogens in milk samples using biocompatible solid-phase microextraction combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). In pretreatments, the biocompatible SPME C18 fiber was conditioned in methanol/water (1/1, v/v) for 10 min before use. The fiber was immersed into 10 mL milk samples for 30 min. After adsorption, the estrogens were desorbed by putting SPME fiber into 1.5 mL acetonitrile for 10min. After derivatized with dansyl chloride, the extract were analyzed by LC- MS/MS. The protein precipitation step is unnecessary in the method proposed. The linearity range was 0.1 to 50 ng/mL . Limits of detection (LODs) of five estrogen were between 6.1 to 14.0 ng/mL. The coefficient of determination were obtained above 0.9956. The relative standard deviations of intra-day and inter-day precisions were 1.5 to 15.7% and 3.9 to 18.2% and the recovery range from 90.1% to 113.9%. The results of this study offered the ng/mL level sensitivity and high selectivity for determination of estrogens in milk. The proposed method is successfully applied to analyze the trace estrogens in milk. The results showed no estrogens was detected in the commercial milk.en_US
dc.description.tableofcontents目錄 謝誌 I 中文摘要 III 英文摘要 IV 目錄 V 表目錄 X 圖目錄 XII 壹、緒論 1 1.1 前言 1 1.2 雌激素介紹 1 1.3 雌激素之檢測分析方法 7 1.4 雌激素衍生化反應 9 1.5 質譜儀原理 11 1.5.1 電灑游離法 13 1.5.2 大氣壓化學游離法 18 1.5.3 串聯質譜儀 20 1.5.4 四極矩質量分析器原理 21 1.6 固相微萃取技術 24 1.6.1固相微萃取法原理 24 1.6.2生物相容性固相微萃取技術 30 1.6.3生物相容性萃取材質 32 1.7 研究動機與目的 35 貳、實驗 37 2.1. 藥品及試劑 37 2.1.1 藥品 37 2.1.2試劑 37 2.2 藥品配製 38 2.2.1 儲存標準溶液 1000 μg/mL 配製 38 2.2.2 標準溶液 10 μg/mL及 1 μg/mL 配製 38 2.2.3 不同濃度混合標準溶液配製 38 2.2.4 碳酸氫鈉緩衝溶液配製 39 2.2.5 衍生化試劑溶液配製 39 2.2.6 含有雌激素之牛奶工作溶液配製 40 2.3 實驗器材與儀器設備 40 2.3.1 實驗器材 40 2.3.2液相層析串聯質譜儀 41 2.3.3 液相層析條件 41 2.3.4 樣品前處理步驟 43 2.3.4.1 生物相容性固相微萃取實驗步驟 43 2.3.4.2 一般固相微萃取實驗步驟 43 2.4 液相層析質譜儀參數之探討 44 2.4.1 液相層析質譜儀離子源之參數探討 44 2.4.2 最佳離子化方法之探討 45 2.4.3 固相微萃取之參數探討 46 參、結果與討論 50 3.1 標準質譜圖之建立 50 3.2 選擇反應偵測之子代離子選擇 51 3.3 液相層析質譜儀離子化參數之探討 65 3.4衍生化於離子化效率之探討 67 3.5 最佳離子化方法之探討 67 3.6 固相微萃取之探討 70 3.6.1 萃取溶液酸鹼度及鹽類添之探討 70 3.6.2 纖維活化時間之探討 72 3.6.3 萃取時間之探討 72 3.6.4 脫附溶劑之探討 74 3.6.5 脫附時間之探討 77 3.7 生物相容性固相微萃取與一般固相微萃取之比較 77 3.8 固相微萃取法分析牛奶中五種雌激素之線性範圍、線性方程式 線性相關係數及偵測極限 80 3.9 固相微萃取分析牛奶中五種雌激素之回收率 83 3.10 固相微萃取分析牛奶中五種雌激素之精確度 83 3.11牛奶真實樣品中雌激素之檢測 86 3.12不同脂肪含量對萃取效率之影響 86 肆、結論 89 伍、參考文獻 90     表目錄 表一、五種雌激素之性質 5 表二、雌激素分析方法文獻 8 表三、固相萃取纖維種類及應用 31 表四、液相層析儀之動相沖提條件 42 表五、電灑游離法負離子模式下,五種未衍生化雌激素之選擇離子斷 裂途徑及其最佳化碰撞能量 58 表六、加熱式電灑游離法正離子模式下,五種雌激素之選擇離子斷裂 途徑和其最佳化碰撞能量 64 表七、正負離子模式下電灑游離法、加熱式電灑游離法及大氣壓化學 游離法偵測雌激素之最佳儀器參數 66 表八、五種雌激素化合物在不同萃取條件下對分析物萃取效率 之影響 71 表九、固相微萃取結合液相串聯質譜術於牛奶中五種雌激素之線性 範圍、線性相關係數、偵測極限 81 表十、固相微萃取結合液相層析串聯質譜術於牛奶中雌激素之 回收率 84 表十一、固相微萃取結合液相層析串聯質譜術於牛奶中雌激素之精確 度評估 85 表十二、固相微萃取結合液相層析串聯質譜術分析牛奶中不同脂肪含 量對雌激素之影響 87   圖目錄 圖一、雌激素主要結構 2 圖二、五種雌激素之結構圖 4 圖三、雌激素於人體中代謝途徑 6 圖四、雌激素與丹磺醯氯之衍生化反應及主要斷裂離子 10 圖五、電灑游離法之電灑現象 14 圖六、電灑游離法之兩種游離化機制 16 圖七、加熱式電灑游離法於正離子模式下離子化過程 17 圖八、大氣壓化學游離法之離子化過程 19 圖九、雙曲面四極矩之結構圖 22 圖十、四極矩質譜儀之結構圖 23 圖十一、商業化固相微萃取裝置圖 26 圖十二、固相微萃取操作流程圖 27 圖十三、生物相容性萃取材質之結構 34 圖十四、商業化生物相容性固相微萃取 C18 纖維 36 圖十五、衍生化後之estriol以電灑游離法正離子模式分析所得之 質譜圖 52 圖十六、衍生化後之estrone以電灑游離法正離子模式分析所得之 質譜圖 53 圖十七、 衍生化後之17α-estradiol以電灑游離法正離子模式分析所得 之質譜圖 54 圖十八、衍生化後之17β-estradiol以電灑游離法正離子模式分析所得之 質譜圖 55 圖十九、衍生化後之17α-ethinylestradiol以電灑游離法正離子模式分析 所得之質譜圖 56 圖二十、電灑游離法正離子模式分析衍生化後estriol 之 [M+H]+, m/z 522所得子代離子質譜圖 59 圖二十一、電灑游離法正離子模式分析衍生化後estrone之 [M+H]+, m/z 504離子所得子代離子質譜圖 60 圖二十二、電灑游離法正離子模式分析衍生化後17α-estradiol 之 [M+H]+, m/z 506離子所得子代離子質譜圖 61 圖二十三、電灑游離法正離子模式分析衍生化後17β-estradiol 之 [M+H]+, m/z 506離子所得子代離子質譜圖 62 圖二十四、電灑游離法正離子模式分析衍生化後17α-ethinylestradiol 之[M+H]+, m/z 530離子所得子代離子質譜圖 63 圖二十五、未衍生化及衍生化對 1 μg/mL 五種雌激素離子化效率之 影響 68 圖二十六、不同游離化方法於正離子模式下對 1 μg/mL 衍生化後 五種雌激素離子化效率之影響 69 圖二十七、不同固相微萃取纖維活化時間對牛奶中 50 ng/mL 五種雌 激素萃取效果之影響 73 圖二十八、不同萃取時間對牛奶中 50 ng/mL 五種雌激素萃取效果之 影響 75 圖二十九、不同脫附溶劑對牛奶中 50 ng/mL 五種雌激素萃取效果之 影響 76 圖三十、不同脫附時間對牛奶中 50 ng/mL 五種雌激素萃取效果之 影響 78 圖三十一、固相微萃取纖維種類對牛奶中 50 ng/mL 五種雌激素萃取 效果之影響 79 圖三十二、添加 50 ng/mL混合標準品於空白脫脂牛奶樣品中,經固 相微萃取結合液相層析串聯質譜術之質譜層析圖 82zh_TW
dc.language.isozh_TWen_US
dc.publisher化學系所zh_TW
dc.relation.urihttp://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-3007201317504600en_US
dc.subject生物相容性固相微萃取zh_TW
dc.subjectEstrogensen_US
dc.subject液相層析串聯質譜zh_TW
dc.subject牛奶zh_TW
dc.subject雌激素zh_TW
dc.subjectMilken_US
dc.subjectBiocompatible Solid-Phase Microextractionen_US
dc.subjectLiquid Chromatography-Tandem Mass Spectrometryen_US
dc.title生物相容性固相微萃取技術結合高效能液相層析串聯質譜術於牛奶中微量雌激素之檢測zh_TW
dc.titleDetermination of Trace Estrogens in Milk Using Biocompatible Solid-Phase Microextraction Combined with Liquid Chromatography-Tandem Mass Spectrometryen_US
dc.typeThesis and Dissertationzh_TW
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