Title page for 91223004


[Back to Results | New Search]

Student Number 91223004
Author Han-Wen Kuo(郭瀚文)
Author's Email Address s1223004@cc.ncu.edu.tw
Statistics This thesis had been viewed 3867 times. Download 4152 times.
Department Chemistry
Year 2003
Semester 2
Degree Master
Type of Document Master's Thesis
Language zh-TW.Big5 Chinese
Title Determination of endocrine disrupting chemicals in different matrices using GC/MS and LC/MS.
Date of Defense 2004-06-25
Page Count 103
Keyword
  • bisphenol A
  • Endocrine disrupters
  • environmental hormones
  • estrogens
  • gas chromatography-mass spectrometry
  • isoflavones
  • liquid chromatography-mass spectrometry
  • phytoestrogen
  • steroid hormones
  • Abstract   Society has been very concerned with the potential risk to human life and wildlife due to exposure to both natural and synthetic estrogenic chemical residues that may interfere with reproduction and development. Although the concentration of these compounds in environmental samples is low, it has been demonstrated that the nonconjugated estrogens at low concentrations can cause the synthesis and secretion of vitellogenin in male fish.
      This study describes a reliable and sensitive method for determining five estrogenic compounds by liquid chromatography/mass spectrometry (LC/MS). Optimization of the ionization process was performed with electrospray ionization, atmospheric pressure chemical ionization and atmospheric pressure photoionization methods using three eluents with different compositions in the LC/MS scan mode, and using both positive ion and negative ion modes. The best ionization sensitivity for the studied compounds was obtained with ESI in negative ion mode when using an acetonitrile-water gradient including 0.1% of ammonium hydroxide (pH 10.7). Using the optimum conditions equipped with reliable pretreatment procedures, the quantitation limit of these compounds was 10 ng/L in 1 L water samples. Recovery of the estrogenic chemicals in spiked various water samples ranged from 77 to 99% while RSD ranged from 1 to 6%.
      Another investigation describes a reliable and sensitive method for simultaneously determining bisphenol A (BPA) and two major phytoestrogens, daidzein and genistein, in powdered milks and infant formulas by gas chromatography–mass spectrometric analysis after trimethylsilylation.
      To reduce the matrix interference associated with the constituents of the formulas, the dissolved formula solutions were firstly ultra-centrifuged and the analytes in the supernatant were then extracted using a C18 solid-phase extraction cartridge. The accuracy and precision of the method were determined and the technique was successfully employed to measure trace concentrations of bisphenol A, daidzein and genistein in powdered formulas.
      The results show that bisphenol A, daidzein and genistein were detected in all the testing samples at concentrations from 45 to 113 ng/g (except one infant formula), 20 to 2050 ng/g and 21 to 6510 ng/g, respectively. The highest concentrations of daidzein and genistein (i.e., 2050 and 6510 ng/g) were detected in a soy-based powdered infant formula. The quantitation limits were 1.0 ng/g for bisphenol A, and 10 ng/g for daidzein and genistein using 0.5 g powdered milk samples.
    Table of Content 中文摘要--------------------------------------------------------------- Ⅰ
    英文摘要--------------------------------------------------------------- Ⅲ
    目錄--------------------------------------------------------------- Ⅴ
    表目錄 ------------------------------------------------------------- Ⅸ
    圖目錄 ------------------------------------------------------------- Ⅹ
    第一章 前言  ---------------------------------------------------------  1
    1-1研究源起  -------------------------------------------------  1
    1-2研究目標  -------------------------------------------------  5
    第二章 文獻回顧  -----------------------------------------------------  8
    2-1環境荷爾蒙  -----------------------------------------------  8
    2-1-1環境荷爾蒙簡介  --------------------------------  8
    2-1-2環境荷爾蒙的分類  ------------------------------  9
         2-2雌性激素類化合物  -----------------------------------------  11
         2-2-1荷爾蒙之簡介  ----------------------------------  11
         2-2-2本研究中雌性激素類化合物之簡介  ----------------  12
         2-2-3水環境中雌性激素類化合物之來源  ----------------  15
         2-2-4國內外分析雌性激素之相關研究  ------------------  16
    2-3植物性雌激素  ---------------------------------------------  20
         2-3-1植物性雌激素對於人體之作用  --------------------  20
         2-3-2植物性雌激素之分類及來源  ----------------------  22
         2-3-3異黃酮素  --------------------------------------  25
         2-3-4國內外分析異黃酮素之相關研究  ------------------  27
    2-4液相層析質譜儀  -------------------------------------------  32
         2-4-1電噴灑游離法  ----------------------------------  33
         2-4-2大氣壓力化學游離法  ----------------------------  34
         2-4-3大氣壓力光游離法  ------------------------------  34
    2-5氣相層析質譜儀  -------------------------------------------  39
         2-5-1四極矩質譜儀  ----------------------------------  39
         2-5-2選擇離子偵測法  --------------------------------  39
    第三章 實驗步驟與樣品分析  -------------------------------------------  41
    3-1實驗藥品與儀器設備  ---------------------------------------  41
         3-1-1實驗藥品  --------------------------------------  41
         3-1-2儀器設備  --------------------------------------  44
    3-2實驗步驟  -------------------------------------------------  45
         3-2-1雌性激素類化合物之分析  ------------------------  45
         3-2-2植物性雌激素與雙酚-A之分析  --------------------  50
    3-3樣品採集  -------------------------------------------------  56
         3-3-1飲用水及地下水樣採集  --------------------------  56
         3-3-2嬰兒奶粉樣品之收集  ----------------------------  56
    第四章 結果與討論  --------------------------------------------------- 57
    4-1雌性激素類化合物之分析  -----------------------------------  57
         4-1-1不同游離源在正負離子模式下之分析結果 -----------  57
         4-1-2不同動相組成溶劑對於分析物感度之影響 -----------  61
         4-1-3雌性激素類化合物之測定  ------------------------  67
         4-1-4檢量線  ----------------------------------------  71
             4-1-5日常用水中的雌性激素類化合物前處理方法探討  ----  73
         4-2植物性雌激素與雙酚-A之分析  -------------------------------  79
         4-2-1衍生化試劑之探討  ------------------------------  79
         4-2-2衍生化溫度與時間的影響  ------------------------  81
         4-2-3異黃酮素與雙酚-A之測定  ------------------------  83
         4-2-4檢量線  ----------------------------------------  86
         4-2-5前處理方法探討  --------------------------------  87
         4-2-6市售嬰兒配方奶粉的分析結果  --------------------  88
    第五章 結論與建議  --------------------------------------------------- 91
         5-1結論  -----------------------------------------------------  91
         5-2建議  -----------------------------------------------------  93
    參考文獻  -------------------------------------------------------------  94
    附錄  -----------------------------------------------------------------  103
    Reference *白珮瑾、何國榮: 2002年諾貝爾化學獎兩種生醫分析的離子化方法,科儀新知,第25卷1期,第7-17頁,民國92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。
    *凌永健: 環境荷爾蒙的化學分析,環境檢驗,第32期,第9-15頁,民國89年。
    *劉朝鑫: 美國FDA對致癌性的動物藥品及飼料添加物的態度,現代畜殖,第15卷1月號,第43-45頁,民國70年。
    *Adlercreutz, H.; Fotsis, T.; Bannwart, C.; Wähälä, K.; Brunow, G.; Hase, T., Isotope dilution gas chromatographic?mass spectrometric method for the determination of lignans and isoflavonoids in human urine, including identification of genistein, Clin. Chim. Acta, 1991, 199, 263-278.
    *Adlercreutz, H.; Fotsis, T.; Kurzer, M.S.; Wähälä, K.; Makela, T.; Hase, T., Isotope dilution gas chromatographic?mass spectrometric method for the determination of unconjugated lignans and isoflavonoids in human feces, with preliminary results in omnivorous and vegetarian women, Anal. Biochem., 1995, 225, 101-108.
    *Anari, M.R.; Bakhtiar, R.; Zhu, B.; Huskey, S.; Franklin, R.B.; Evans, D.C., Derivatization of ethinylestradiol with dansyl chloride to enhance electrospray ionization: application in trace analysis of ethinylestradiol in rhesus monkey plasma, Anal. Chem., 2002, 74, 4136-4144.
    *Ardrey, R.E., Liquid chromatography–mass spectrometry: an introduction, John Wiley & Sons, Ltd., 2003.
    *Avery, M.J., Quantitative characterization of differential ion suppressionn on liquid chromatography/atmospheric pressure ionization mass spectrometric bioanalytical methods, Rapid Commun. Mass Spectrom., 2003, 17, 197-201.
    *Barnes, K.A.; Smith, R.A.; Williams, K.; Damant, A.P.; Shepherd, M.J., A microbore high performance liquid chromatography/ electrospray ionization mass spectrometry method for the determination of the phytoestrogens genistein and daidzein in comminuted baby foods and soya flour, Rapid Commun. Mass Spectrom., 1998, 12, 130-138.
    *Beekman, M.C.; Lingeman, H.; Brinkman, U.A.T.; Gooijer, C., Determination of the isoflavone formononetin in red clover (Trifolium pratense L.) by micellar electrokinetic chromatography combined with deep-UV laser-induced wavelength-resolved fluorescence detection, J. Microcolumn Separations, 1999, 11(5), 347-352.
    *Benijts, T.; Dams, R.; Günther, W.; Lambert, W.; de Leenheer, A., Analysis of estrogenic contaminants in river water using liquid chromatography coupled to ion trap based mass spectrometry, Rapid Commun. Mass Spectrom., 2002, 16, 1358-1364.
    *Benijts, T.; Lambert, W.; de Leenheer, A., Analysis of multiple endocrine disruptors in environmental waters via wide-spectrum solid-phase extraction and dual-polarity ionization LC-ion trap- MS/MS, Anal. Chem., 2004, 76, 704-711.
    *Bennets, H.W.; Underwood, E.J.; Sheir, F.L., A specific breeding problems of sheep on subterranean clover pastures in Western Australia, Australian Veterinary Journal, 1946, 22, 2-5.
    *Borgert, C.J.; LaKind, J.S.; Witorsch, R.J., A critical review of methods for comparing estrogenic activity of endogenous and exogenous chemicals in human milk and infant formula, Environ. Health Perspect., 2003, 111(8), 1020-1036.
    *Bruins, A.P., Mechanistic aspects of electrospray ionization, J. Chromatogr. A, 1998, 794, 345-357.
    *Chen, G.; Zhang, H.W.; Ye, J.N., Determination of rutin and quercetin in plants by capillary electrophoresis with electrochemical detection, Anal. Chim. Acta, 2000, 423, 69-76.
    *Chen, G.; Zhang, J.X.; Ye, J.N., Determination of puerarin, daidzein and rutin in Pueraria lobata (Wild.) Ohwi by capillary electrophoresis with electrochemical, J. Chromatogr. A, 2001, 923, 255-262.
    *Croley, T.R.; Hughes, R.J.; Koenig, B.G.; Metcalfe, C.D.; March, R.E., Mass spectrometry applied to the analysis of estrogens in the environment, Rapid Commun. Mass Spectrom., 2000, 14, 1087-1093.
    *Díaz-Cruz, M.S.; López de Alda, M.J.; López, R.; Barceló, D., Determination of estrogens and progestogens by mass spectrometric techniques (GC/MS, LC/MS and LC/MS/MS), J. Mass Spectrom., 2003, 38, 917-923.
    *Ding, W.H.; Chiang, C.C., Derivatization procedures for the detection of estrogenic chemicals by gas chromatography/mass spectrometry, Rapid Commun. Mass Spectrom., 2003, 17, 56-63.
    *de Rijke, E.; Zafra-Gomez, A.; Ariese, F.; Brinkman, U.A.Th.; Gooijer, C., Determination of isoflavone glucoside malonates in Trifolium pratense L. (red clover) extracts: quantification and stability studies, J. Chromatogr. A, 2001, 932, 55-64.
    *Ferguson, P.L.; Iden, C.R.; McElroy, A.E.; Brownawell, B.J., Determination of steroid estrogens in wastewater by immunoaffinity extraction coupled with HPLC–Electrospray-MS, Anal. Chem., 2001, 73, 3890-3895.
    *Franke, A.A.; Custer, L.J., Daidzein and genistein concentrations in human milk after soy consumption, Clinical Chemistry, 1996, 42(6), 955-964.
    *Fukazawa, H.; Hoshino, K.; Shiozawa, T.; Matsushita, H.; Terao, Y., Identification and quantification of chlorinated bisphenol A in wastewater from wastepaper recycling plants, Chemosphere, 2001, 44, 973-979.
    *Gangl, E.T.; Annan, M.; Spooner, N.; Vouros, P., Reduction of signal suppression effects in ESI-MS using a nanosplitting device, Anal. Chem., 2001, 73, 5635-5644.
    *Garrett, S.D.; Lee, H.A.; Friar, P.M.K.; Morgan, M.R.A., Validation of a novel estrogen receptor-based microtitrationplate assay for the determination of phytoestrogens in soy-based foods, J. Agric. Food Chem., 1999, 47, 4106-4111.
    *Griffith, P.; Collison, M.W., Improved methods for the extraction and analysis of isoflavones from soy-containing foods and nutritional supplements by reversed-phase high-performance liquid chromatography and liquid chromatography–mass spectrometry, J. Chromatogr. A, 2001, 913, 397-413.
    *Hansen, P.D.; Dizer, H.; Hock, B.; Marx, A.; Sherry, J.; McMaster, M.; Blaise, Ch., Vitellogenin - a biomarker for endocrine disruptors, Trends Anal. Chem., 1998, 17, 448-451.
    *Hutabarat, L.S.; Greenfield, H.; Mullholland, M., Quantitative determination of isoflavones and coumestrol in soybean by column liquid chromatography, J. Chromatogr. A, 2000, 886, 55-63.
    *Ibarreta, D.; Daxenberger, A.; Meyer, H.H.D., Possible health impact of phytoestrogens and xenoestrogens in food, APMIS, 2001, 109, 161-184.
    *Ingrand, V.; Herry, G.; Beausse, J.; de Roubin, Marie-Renée, Analysis of steroid hormones in effluents of wastewater treatment plants by liquid chromatography–tandem mass spectrometry, J. Chromatogr. A, 2003, 1020, 99-104.
    *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.
    *Kelly, C., 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, 2000, 872, 309-314.
    *Knight, D.C.; Eden, J.A., A review of the clinical effects of phytoestrogens, Obset Gynecol., 1996, 87(5 Pt 2), 897-904.
    *Lee, H.B.; Peart, T.E., Residues and trace elements - determination of 17?-estradiol and its metabolites in sewage effluent by solid-phase extraction and gas chromatography/mass spectrometry, J. AOAC Int., 1998, 81(6), 1209-1216.
    *Li, D.; Park, J.; Oh, J.R., Silyl derivatization of alkylphenols, chlorophenols, and bisphenol A for simultaneous GC/MS determination, Anal. Chem., 2001, 73, 3089-3095.
    *López de Alda, M.J.; Barceló, D., Determination of steroid sex hormones and related synthetic compounds considered as endocrine disrupters in water by fully automated on-line solid-phase extraction–liquid chromatography–diode array detection, J. Chromatogr. A, 2001, 911, 203-210. (a)
    *López de Alda, M.J.; Barceló, D., Use of solid-phase extraction in various of its modalities for sample preparation in the determination of estrogens and progestogens in sediment and water, J. Chromatogr. A, 2001, 938, 145-153. (b)
    *Mallet, C.R.; Lu, Z.; Mazzeo, J.R., A study of ion suppression effects in electrospray ionization from mobile phase additives and solid-phase extracts, Rapid Commun. Mass Spectrom., 2004, 18, 49-58.
    *Mouatassim-Souali, A.; Tamisier-Karolak, S.L.; Perdiz, D.; Cargouet, M.; Levi, Y., Validation of a quantitative assay using GC/MS for trace determination of free and conjugated estrogens in environmental water samples, J. Sep. Sci., 2003, 26, 105-111.
    *Ososki, A.L.; Kennelly, E.J., Phytoestrogen: a review of the present state of research, Phytother. Res., 2003, 17, 845-869.
    *Perrin, C.; Meyer, L.; Mujahid, C.; Blake, C.J., The analysis of 5′- mononucleotides in infant formulae by HPLC, Food Chem., 2001, 74, 245-253.
    *Petrovic, M.; Tavazzi, S.; Barcelo, D., Column-switching system with restricted access pre-column packing for an integrated sample cleanup and liquid chromatographic–mass spectrometric analysis of alkylphenolic compounds and steroid sex hormones in sediment, J. Chromatogr. A, 2002, 971, 37-45.
    *Pinnella, K.D.; Cranmer, B.K.; Tessari, J.D.; Cosma, G.N.; Rao Veeramachaneni, D.N., Gas chromatographic determination of catecholestrogens following isolation by solid-phase extraction, J. Chromatogr. B, 2001, 758, 145-152.
    *Purdom, C.E.; Hardiman, P.A.; Bye, V.J.; Eno, N.C.; Tyler, C.R.; Sumpter, J.P., Estrogenic effects of the effluent from sewage treatment works, Chem. Ecol., 1994, 8, 275-285.
    *Raffaelli A.; Saba, A., Atmospheric pressure photoionization mass spectrometry, Mass Spec. Rev., 2003, 22, 318-331.
    *Robb, D.B.; Covey, T.R.; Bruins, A.P., Atmospheric pressure photoionization: an ionization method for liquid chromatography– mass spectrometry, Anal. Chem., 2000, 72, 3653-3659.
    *Satterfield, M.; Black, D.M.; Brodbelt, J.S., Detection of the isoflavone aglycones genistein and daidzein in urine using solid-phase microextraction/high-performance liquid chromatography/electrospray ionization mass spectrometry, J. Chromatogr. B, 2001, 759, 33-41.
    *Seifert, M.; Brenner-Weiß, G.; Haindl, S.; Nusser, M.; Obst, U.; Hock, B., A new concept for the bioeffects-related analysis of xenoestrogens: Hyphenation of receptor assays with LC-MS, Fresenius J. Anal. Chem., 1999, 363, 767-770.
    *Setchell, K.D.R.; Zimmer-Nechemias, L.; Cai, J.; Heubi, J.E., Exposure of infants to phyto-oestrogens from soy-based infant formula, Lancet, 1997, 350, 23-27.
    *Setchell, K.D.R.; Zimmer-Nechemias, L.; Cai, J.; Heubi, J.E., Isoflavone content of infant formulas and the metabolic fate of these phytoestrogens in early life, Am. J. Clin. Nutr., 1998, 68, 1453-1461.
    *Setchell, K.D.R.; Cassidy, A., Dietary isoflavones: biological effects and relevance to human health, J. Nutr., 1999, 129(3), 758-767.
    *Sheen, J.F.; Her, G.R., Analysis of hydroxyl-drugs in human  plasma by fluoride attachment in negative ion electrospray   liquid chromatography–mass spectrometry, International Chemical Conference, Taipei: Analytical Chemistry, 2003, P061.
    *Smets, F.; Vanhoenackere, Ch.; Pottie, G., Influence of matrix and applied method on the detection of anabolic residues in biological samples, Anal. Chim. Acta, 1993, 275, 147-162.
    *Svenson, A.; Allard, Ann-Sofie; Ek, M., Removal of estrogenicity in Swedish municipal sewage treatment plants, Water Research, 2003, 37, 4433-4443.
    *Tekel, J.; Daeseleire, E.; Heeremans, A.; van Peteghem, C., Development of a simple method for the determination of genistein, daidzein, biochanin A, and formononetin (biochanin B) in human urine, J. Agric. Food Chem., 1999, 47, 3489-3494.
    *Ternes, T.A.; Andersen, H.; Gilberg, D.; Bonerz, M., Determination of estrogens in sludge and sediments by liquid extraction and GC/MS/MS, Anal. Chem., 2002, 74, 3498-3504.
    *Vanderford, B.J.; Pearson, R.A.; Rexing, D.J.; Snyder, S.A., Analysis of endocrine disruptors, pharmaceuticals, and personal care products in water using liquid chromatography/tandem mass spectrometry, Anal. Chem., 2003, 75, 6265-6274.
    *Wang, C.C.; Prasain, J.K.; Barnes, S., Review of the methods in the determination of phytoestrogens, J. Chromatogr. B, 2002, 777, 3-28.
    *Wang, C.Y.; Huang, H.Y.; Kuo, K.L.; Hsieh, Y.Z., Analysis of Puerariae radix and its medicinal preparations by capillary electrophoresis, J. Chromatogr. A, 1998, 802, 225-231.
    *Zavitsanos, P.; Miller, C., Identification of steroids in water by ion trap LC/MS/MS, Agilent Technologies publication, 2002, 5988-6926EN.
    Advisor
  • Wang-Hsien Ding(丁望賢)
  • Files
  • 91223004.pdf
  • approve immediately
    Date of Submission 2004-06-28

    [Back to Results | New Search]


    Browse | Search All Available ETDs

    If you have dissertation-related questions, please contact with the NCU library extension service section.
    Our service phone is (03)422-7151 Ext. 57407,E-mail is also welcomed.