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研究生： 鍾裕霖
Yu-Lin Chung
論文名稱： 非水相毛細管電泳/低溫螢光分析法對尿液中3,4-亞甲雙氧甲基安非他命(3,4-MDMA)之分析研究
On-line Identification of 3,4-methylenedioxymethamphetamine in Human Urine by Nonaqueous Capillary Electrophoresis Fluorescence Spectroscopy at 77 K
指導教授： 林震煌
Lin, Cheng-Huang
學位類別： 碩士
Master
系所名稱： 化學系
Department of Chemistry
論文出版年： 2001
畢業學年度： 89
語文別： 中文
論文頁數： 146
中文關鍵詞： 非水相 、毛細管電泳 、3,4-亞甲雙氧甲基安非他命 、螢光光譜 、77 K
英文關鍵詞： nonaqueous, capillary electrophoresis, 3,4-MDMA, fluorescence spectroscopy, 77 K
論文種類： 學術論文
相關次數： 點閱：290  下載：0 
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    • 3,4-MDMA（3,4-methylenedioxymethamphetamine）俗稱快樂丸，服用之後會導致腦內關於思考與記憶的功能長期下降，甚至可能會損害腦的永久功能，現已經成為國內最常見的濫用藥物。文獻記載MDMA與其異構物的鑑定法，大多使用GC-MS，必要時輔以衍生化的反應來協助分析。然而，此類化合物結構類似，尤其是經體內代謝過後的物質，不容易單由質譜的斷裂譜峰來加以判別。為了能快速且可正確判知MDMA類化合物的種類，本研究首創非水相毛細管電泳層析/超低溫螢光分析法，先以電泳的方式分離樣品，當待測物到達螢光偵測窗時，瞬間倒入液態氮，將毛細管冷卻到極低（77 K）的溫度。由於MDMA在低溫之下光譜的頻寬要比室溫時更窄，而且可以觀測到明顯的振電躍遷譜線，因此有利於光譜之指紋比對。本研究更發現以非水相螢光偵測的方法，相對於一般水相緩衝溶液的分離法，可使樣品螢光強度不受水相溶液的削弱影響，當使用SC 100 mM、ammonium acetate 20 mM，溶於formamide/methanol/ethanol（30:20:50 v/v/v）所配製成的緩衝溶液來分離，每次分離所需時間僅約10分鐘，即可將分析物與尿液中的干擾物分開。本研究所開發的方法其偵測極限為1 ppm（5.2 ´ 10-6 M），加上使用簡單的液-液萃取法，可使偵測極限降至150 ppb。本研究使用憲兵司令部刑事鑑識中心提供的標準品與不同濫用藥物者的尿液，經由低溫光譜比對能有效鑑定MDMA吸毒者尿液中的MDMA含量。再者，本研究獨創非水相冰溫分離法，將1/3長的毛細管浸於零下50度的乾冰槽中的分離方式，相對於一般在室溫之下進行分離的方法，可以提高電泳分離的解析度2.5倍！

    We have demonstrated that capillary electrophoresis (CE) can be easily interfaced with 77 K luminescence spectroscopy (LS) for separation and on-line spectral identification of structurally similar analytes. This novel CE-LS apparatus consists of a regular CE system, instrumentation for luminescence spectroscopy, and a specially designed capillary-dewar. When the separating molecules traverse into the cryostat detection window liquid nitrogen is added, freezing the separating analyte zones within the capillary. We present the first application of the CE-LS system to structural isomers (2, 3- and 3, 4-methylenedioxymethamphetamine). Then application of this system for 3,4-MDMA and related amphetamines in urine samples are described for nonaqueous capillary electrophoresis/fluorescence spectroscopy and identified on-line at 77 K. Under optimized conditions, baseline separation of the selected compounds was achieved in less than 12 min. Precision was evaluated by measuring the repeatability and intermediate precision of the migration times and corrected peak areas. The nonaqueous CE separation conditions and the spectral characteristics of 3,4-MDMA with respect to solvent and temperature effects are also discussed.

    中文摘要 .........................................Ⅰ 英文摘要 .........................................Ⅱ 目錄 .........................................Ⅲ 圖目錄 .........................................Ⅵ 表目錄 .........................................Ⅷ 第一章、緒論.......................................1 1-1.分析物簡介.....................................1 1-1-1. 安非他命的性質.....................1 1-1-2. 3,4-MDMA..........................3 1-1-3. MDMA類濫用藥物....................6 1-2.研究目的.......................................8 第二章、毛細管電泳 ................................11 2-1 毛細管電泳的發展緣由 .......................11 2-2 毛細管電泳的基本原理 .......................15 2-2-1. 電泳的分離與遷移率 .......................15 2-2-2. Stern的電雙層理論 .......................16 2-2-3. 管柱分離效率 .......................19 2-2-4. 常見的毛細管電泳分離模式 ..............20 2-2-4-1. 毛細管區帶電泳（CZE） ..............20 2-2-4-2. 微胞電動毛細管層析（MEKC） ..............23 2-3 非水相毛細管電泳法 .......................26 2-4 低溫（77 K）螢光光譜分析法 ..............28 2-5 非水相毛細管電泳/低溫（77 K）螢光分析法 .....32 第三章、儀器及藥品 ............................... 35 3-1 藥品 ..........................................35 3-2 儀器及其週邊設備 ........................38 3-3 Xe燈光源波長的選擇及功率的測量 ...............41 3-4 自組式毛細管電泳螢光分析儀 ...............45 3-5 Off-line螢光光譜的測量 ........................47 3-6 毛細管電泳層析分離／低溫（77 K）螢光分析法......49 3-7 毛細管電泳冰溫層析分離法的開發 ...............52 3-8 雷射激發螢光／螢光lifetime偵測系統..............55 第四章、實驗結果與討論...............................57 4-1 MDMA化合物的光譜性質 .........................57 4-1-1. 吸收光譜 ..................................57 4-1-2. 室溫與77 K之下的螢光光譜與螢光激發光譜 .......63 4-1-3. 3,4-MDMA的磷光性質與重原子效應...............68 4-1-4. 3,4-MDMA在77 K之下的磷光與螢光壽命...........71 4-1-5. 溶劑對低溫螢光光譜的影響......................75 4-1-6. MDMA類化合物螢光光譜之比較...................77 4-2 On-line 低溫螢光光譜之鑑定......................80 4-2-1. 分離條件的確立─非水相分離....................80 4-2-2. 非水相CE緩衝的探討 ..........................84 4-2-2-1. 黏度 ...................................84 4-2-2-2. 樣品注入量..................................86 4-2-2-3. 介電常數 ...................................87 4-2-2-4. pH* ...................................87 4-2-2-5. 界面活性劑SC的性質 ..........................89 4-2-2-6. CMC*的測量..................................91 4-2-2-7. 分離順序反轉.................................94 4-2-2-8. 分離的最佳化條件.............................96 4-3 3,4-MDMA的檢量線.................................98 4-4 2,3-MDMA及3,4-MDMA的分離與on-line 77 K光譜......102 4-5 吸毒者尿液中3,4-MDMA的定性及定量 ................106 4-5-1. 尿液樣品的保存與前處理 ................106 4-5-2. 尿液的萃取 ...................................106 4-5-3. 萃取回收率 ....................................108 4-5-4. 吸毒者尿液萃取的分析及On-line 77 K光譜指紋比對..110 4-6 溫度對異構物分離的影響 ...........................113 第五章、結論...........................................117 發表論文 ..............................................120 參考文獻 ..............................................121 附錄...................................................127 附錄1 管制藥品的分級...................................127 附錄2 管制藥品登記證...................................134 附錄3 3,4-MDMA及其他類似化合物之光譜性質..............135 附錄3-1 3,4-MDMA及3,4-BDB的質譜.......................135 附錄3-2 3,4-MDMA之FT-IR光譜...........................136 附錄3-3 3,4-MDMA及其他類似化合物之拉曼光譜............137 附錄3-4 其他MDMA衍生物之螢光光譜.......................138 附錄4 毒品濫用之尿液檢定標準...........................144

    1. J. L. Cadet, S.V. Ordonez, and J. V. Ordonez, Synapse 25:176-184, 1997.
    2. X. Deng, B. Ladenheim, L. I. Tsao, and J. L. Cadet, Journal of Neuroscience 19(22):10107-10115, 1999.
    3. W. P. Melega, Brain Research, in press.
    4. U. D. McCann, D. F. Wong, F. Yokoi, V. Villemagne, R. F. Dannals, and G. A. Ricaurte, Journal of Neuroscience 18(20):8417-8422, 1998.
    5. NIDA’s Home Page, http://www.nida.nih.gov/Infofax/ecstasy.html
    6. NIDA’s Home Page, Some facts about club drugs, Jaunary 25, 2001. http://165.112.78.61/ClubAlert/ClubDrugAlert.html
    7. NIDA’s Home Page, Facts About MDMA (Ecstasy), NIDA Notes, Vol 14, N 4, http://165.112.78.61/NIDA_Notes/NNVol14N4/tearoff.html
    8. S. J. Peroutka, N Engl J Med 1987, 317, 1542-1543.
    9. K. Verebey, J. Alrazi, J. H. Jaffe, JAMA(R) 1988, 259, 1649-1650
    10. 管制藥品局/預警宣導組/常見濫用物質及其毒害/苯二氮泮類安眠鎮定劑 http://www.nnb.gov.tw/prop/prop_1-08.asp
    11. 台北市衛生局 http://www.health.gov.tw/News.asp?News=495&Key=685
    12. Fischer, C.; Hatzidimitriou, G.; Wlos, J.; Katz, J.; and Ricaurte, G. Journal of Neuroscience 15 :5476- 5485, 1995.
    13. Bolla, K.I.; McCann, U.D.; and Ricaurte, Neurology 51 :1532-1537, 1998.
    14. Hatzidimitriou, G.; McCann, U.D.; and Ricuarte, Journal of Neuroscience 191(12):5096-5107, 1999.
    15. McCann, U.D.; Mertl, M.; Eligulashvili, V.; and Ricaurte, Psychopharmacology 143:417-425, 1999.
    16. McCann, U.D.; Szabo, Z.; Scheffel, U.; Dannals, R.F.; and Ricaurte, G.A. Positron emission tomographic evidence of toxic effect of MDMA ("ecstasy") on brain serotonin neurons in human beings.
    17. http://ibbserver.ibb.uu.nl/~jboschma/ecstasy/xtc01
    18. J. T. Cody, R. Schwarzhoff, J. Anal. Toxicol. 17 (1993) 26.
    19. R. J. Renton; J. S. Cowie, M. C. Oon, Forensic Science International, 60 (1993 Aug), 189
    20. O. Beck, M. Kraft, M. R. Moeller, B. L. Smith, S. Schneider, R. Wennig, Ann. Clin. Biochem. 37 (2000) 199.
    21. K. M. Clauwaert, J. F. Van Bocxlaer, E. A. De Letter, S. Van Calenbergh, W. E. Lambert, A. P. De Leenheer, Clin. Chem. (Washington, D. C.) 46 (2000) 1968.
    22. F. Tagliaro, Z. De Battisti, A. Groppi, Y. Nakahara, D. Scarcella, R. Valentini, M. Marigo, J. Chromatogr., B: Biomed. Sci. Appl. 723 (1999) 195.
    23. R. E. Michel, A. B. Rege, W. J. George, J. Neurosci. Methods 50 (1993) 61.
    24. S. Borth, W. Hansel, P. Rosner, T. Junge, J. Mass Spectrom. 35 (2000) 705.
    25. J. L. Valentine, R. Middleton, J. Anal. Toxicol. 24 (2000) 211.
    26. M. Guidotti, G. Ravaioli, Boll. Chim. Ig., Parte Sci. 50 (1999) 213.
    27. C. Jurado, M. P. Gimenez, T. Soriano, M. Menendez, M. Repetto, J. Anal. Toxicol. 24 (2000) 11.
    28. D. Talwar, I. D. Watson, M. J. Stewart, J. Chromatogr., B: Biomed. Sci. Appl. 735 (1999) 229.
    29. D. Hensley, J. T. Cody, J. Anal. Toxicol. 23 (1999) 518.
    30. O. A. Silva, M. Yonamine, V. E. D. Reinhardt,. Rev. Farm. Bioquim. Univ. Sao Paulo. 34 (1998) 33.
    31. Y. Nakahara, R. Kikura, Biol. Pharm. Bull. 20 (1997) 969.
    32. U. Staerk,W. R. Kulpmann, J. Chromatogr., B: Biomed. Sci. Appl. 745 (2000) 399.
    33. W. Lu, G. K. Poon, P. L. Carmichael, R. B. Cole, Anal. Chem. 68 (1996) 668.
    34. H. Qzaki, N. Itou, N.; S. Terabe, T. Takada, M. Sakairi, H. Koizumi, J. Chromatogr. A 716 (1995) 69.
    35. J. Cai, J. Henion, J. Anal. Toxicol. 20 (1996) 27.
    36. S. J. Locke, P. Thibault, Anal. Chem. 66 (1994) 3436.
    37. J. Cai, J. Henion, J. Chromatogr. A. 703 (1995) 667.
    38. D. L. Olson, M. E. Lacey, J. V. Sweedler, Anal. Chem. 70 (1998) 645.
    39. R. Todorowic, F. Ariese, P. Devanesan, R. Jankowiak, G. J. Small, E. G. Rogan, E. L. Cavalieri, Chem. Res. Toxicol., 10 (1997) 941.
    40. D. Zamzow, C. -H. Lin, G. J. Small, R. Jankowiak, J. Chromatogr. A 781 (1997) 73.
    41. M. Shu, F. Ariese, G. J. Small, R. Jankowiak, A. Hewer, D. H. Phillips, Carcinogenesis, 16 (1995) 2561.
    42. R. Jankowiak, D. Zamzow, W. Ding, G. J. Small, Anal. Chem. 68 (1996) 2549.
    43. K. P. Roberts, C. -H. Lin, R. Jankowiak, G. J. Small, J. Chromatogr. A 853 (1999) 159.
    44. R. Jankowiak, K. P. Roberts, G. J. Small, Electrophoresis 21 (2000) 1251.
    45. K. P. Roberts, C. -H. Lin, M. Singhal, G. P. Casale, G. J. Small, R. Jankowiak,. Electrophoresis 21 (2000) 799.
    46. R. Jankowiak, C. -H. Lin, D. Zamzow, K. P. Roberts, K. -M. Lim G. J. Small, Chem. Res. Toxicol. 12 (1999) 768.
    47. F. Kohlrausch, Wiedemanns Ann. Phys. Chem., 62(1897) 209-239.
    48. S. Hjerten, Chromatogr. Rev., 9(1967) 122-239.
    49. P. Jandik and G. Bonn, Capillary Electrophoresis of Small Molecules and Ions, VCH, New York, 1993.
    50. S. Terabe, K. Otsuka, K. Ichikawa, A. Tsuchiya and T. Ando, Anal. Chem., 56 (1984), 111.
    51. S. Terabe, K. Otsuka, and T. Ando, Anal. Chem., 1985, 57, 834.
    52. K. H. Row, W. H. Griest, M. P. Maskarienc, J.Chromatogr., 409(1987) 193.
    53. S. F. Y. Li, Capillary electrophoresis, 1992, chapter 1, pp 1-5.
    54. S. Compton, R. Brownlee, Biotechniques, 1988, 6, 432
    55. W. B. Hardy, J. Physiol., 1892, 24, 288.
    56. W. B. Hardy, J. Physiol., 1905, 33, 273.
    57. T. B. Coolidge, J. Biol. Chem., 1939, 127,551.
    58. R. A. Consden, A. H. Gorden, and A. J. P. Martin, Biochem. J., 1946, 40, 33.
    59. J. Porath, Biochem. Biophys. Acta., 1956, 22, 151.
    60. S. Hjerten, Chromatogr. Rev., 1967, 9, 122.
    61. F. E. P. Mikkers, F. M. Everaerts, and T. P. E. M. Verheggen, J. Chromatogr., 1979, 169, 11.
    62. A. Tiselius, Trans. Faraday. Soc., 33(1997) 524.
    63. J. Knox, Chromatographia, 26(1988) 329.
    64. R. Virtanen, Acta Polytechnica Scand., 123(1979) 1-67.
    65. J. W. Joegenson and K. D. Lukacs, J. Chromatogr., 218(1981) 209-216.
    66. J. W. Joegenson and K. D. Lukacs, Anal. Chem., 53(1981) 1298-1302.
    67. D. J. Rose and J. W. Jorgenson, Anal. Chem., 1988, 60, 642.
    68. S. Hjerten and M. D. Zhu, J.Chromatogr., 346(1985) 265.
    69. S. Hjertén, J. L. Liao, and K. Yao, J. Chromatogr., 1987, 387, 127.
    70. A. Cohen and B. L. Karger, J. Chromatogr., 397(1987) 409.
    71. X. Huang and R. N. Zare, Anal. Chem., 63(1991) 2193.
    72. X. Huang, M. J. Gordon, and R. N. Zare, Anal. Chem., 60(1988) 375.
    73. R. D. Holland and M. J. Sepaniak, Anal. Chem., 65(1993) 1140.
    74. R. T. Kennedy and J. W. Jorgenson, Anal. Chem., 61(1989) 1128.
    75. W. Lu, G. K. Poon, P. L. Carmichael, R. B. Cole, Anal. Chem. 68 (1996) 668-674.
    76. C. -H. Lin, Y. -L. Chung, Y-H. Chen, Analyst, 126 (2001) 302-305
    77. Y. –L. Chung, J. –T. Liu, and C. –H. Lin, J. Chromatogr. B, 2001, in press.
    78. R. J. Hunter, Zeta potential in Colloid Science: Principles and Applications, Academic Press, New York, 1981, Chap 2.
    79. High Performance Capillary Electrophoresis - An Introduction, Dr. David N. Heiger, 1992, 2nd edition.
    80. H. Z. Helmholtz, Anal. Phys. Chem., 1897, 7, 337.
    81. B. Krattiger, G. J. M. Bruin, and A. E. Bruin, Anal. Chem. 66 (1994) 1.
    82. M. Stefansson and M. Novotny, Anal. Chem. 66 (1994) 1134.
    83. Y. Kim and M. D. Morris, Anal. Chem. 66 (1994) 1168.
    84. Z. Zhao, A. Malik, and M. L. Lee, Anal. Chem. 65 (1994) 2747.
    85. M. Hayek, J. Phys. Colloid. Chem. 55 (1951) 1527-1533.
    86. M. M. Tuckerman, H. H. Strain, Anal. Chem. 32 (1960) 695-698.
    87. N. J. Parekh, A. A. Fatmi, M. A. Tshabalala, B. L. Rodgers, J. Chromatogr. 314 (1984) 65-82.
    88. S. Fujiwara, S. Honda, Anal. Chem. 59 (1987) 487-490.
    89. A. J. Tomlinson, L. M. Benson, J. P. Landers, G. F. Scanlan, J. Fang, J. W. Gorrod, S. J. Naylor, J. Chromatogr. 652 (1993) 417-426.
    90. A. J. Tomlinson, A. J. Tomlinson, J. M. Reid, D. L. Walker, M. M. Ames, S. J. Naylor, High Resolut. Chromatogr. 16 (1993) 324-326.
    91. A. J. Tomlinson, A. J. Tomlinson, S. J. Naylor, High Resolut. Chromatogr. 176 (1994) 175-177.
    92. Y. Walbroehl, J.W. Jorgenson, J. Chromatogr. 315 (1984) 135-143.
    93. R. Carabias-Martý´nez,* E. Rodrý´guez-Gonzalo, J. Domý´nguez-Alvarez, and J. Herna´ ndez-Me´ ndez, Anal. Chem. 69 (1997) 4437-4444
    94. D. L. Gallaher, J. and M. E. Johnson, Anal. Chem. 72 (2000) 2080-2086.
    95. V. L. Ward, M. G. Khaledi, J. Chromatogr. A. 859 (1999) 203-219.
    96. F. Wang, M. G. Khaledi, Anal. Chem. 68 (1996) 3460-3467.
    97. F. Wang, M. G. Khaledi, J. Chromatogr. A, 817 (1998) 121-128.
    98. F. Wang, M. G. Khaledi, J. Chromatogr. A, 875 (2000) 277-293.
    99. V. L. Ward, M. G. Khaledi, J. Chromatogr. B, 718 (1998) 15-22.
    100. A. J. Tomlinson, L. M. Benson, S. Naylor, LC-GC 8 (1995) 210-216.
    101. E. V. Shpol’skii, A. A. Il’ina, and L. A. Klimova, Dokl. Akad. Nauk SSSR 87, 935 (1952).
    102. Zhe Jin, Rong Chen, and Luis A. Colón* ; Analytical Chemistry; 1997; 69(7); 1326-1331
    103. Separation of carnitine and acylcarnitines by capillary electrophoresis. Journal of Chromatography A, Volume: 781, Issue: 1-2, September 26, 1997, 475-479
    104. Enzyme immunoassay by capillary electrophoresis with laser induced fluorescence detection. Analytica Chimica Acta, Volume: 399, Issue: 1-2, November 8, 1999, 63-68.
    105. Trace detection of NO and NO2 by photoionization and laser induced fluorescence techniques. Analytica Chimica Acta, Volume: 397, Issue: 1-3, October 4, 1999, 3-41.
    106. A. M. Phipps, D. N. Hume, J. Chem. Educ., 1968, 45, 664.
    107. V. Vorsa, K. F. Willey, N. Winograd,. Anal. Chem. 1999 71, 574-581
    108. McCLURE, J. Chem. Phys., 17 905 (1949).
    109. KASHA, J. Chem. Phys., 20 71 (1952).
    110. P. W. Atkins, in: Physical Chemistry, 4th ed., W. H. Freeman, New York, 1990, Ch. 24.
    111. D. S. Burgi, K. Salomon, R. L. Chien, J. Liq. Chromatogr. 14 (1991) 847-867.
    112. M. L. Riekkola, S. K. Wiedmer, I. E. Valkó, H. Sirén, J. Chromatogr. A, 792 (1997) 13-35.
    113. K. Otsuka, S. Terabe, J. Chromatogr. A, 875 (2000) 163-178.
    114. H. Sirén, T. Hiissa, Y. Min, Analyst, 2000 (125) 1561-1568.
    115. H. Nishi, J. Chromatogr. A, 735 (1996) 57-76.
    116. A. N. Díaz, F. G. Sánchez, A. G. Pareja, Colloids and Surface A: Physicochemical and Engineering aspects. 142 (1998) 27-34
    117. A. Cifuentes, J. L. Bernal, J. C. Diez-Masa, Anal. Chem. 69 (1997) 4271-4274.
    118. A. Amini, I. Beijersten, C. Pettersson, D. Westerlund, J. Chromatogr. A 737 (1996) 301–313.
    119. U. Pyell, U. Bütehorn, Chromatographia, Vol. 40, No. 1/2, Janurary 1995.
    120. S. Ma, F. Kálmán, A. Kálmán, F. Thunecke, C. Horváth, J. Chromatogr. A, 716 (1995) 167-182.
    121. C. M. Park, Ann. NY Acad. Sci. 209 (1973) 237
    122. M. Perrella, A. Heyda, A. Mosca, L. Rossi-Bernardi, Anal. Biochem. 88 (1978) 212.
    123. T. Komuro, C. Yomota, H. Isaka, Chem. Pharm. Bull. 36 (1988) 1218.
    124. S. Ma, Cs. Horváth, Electrophoresis 18 (1997) 873.
    125. S. Ma, Cs. Horváth, J. Chromatogr. A, 825 (1998) 55-69.
    126. F. T. Noggle, C. R. Clark, J. D. Ruiter, J. Liq. Chromatog., 14(10), 1991, 1913.
    127. Steven E. J. Bell, D. T. Burns, C. Dennis, James S. Speers, Analyst, 2000, 125, 541–544.
    128. K. Ueno, and E. S. Yeung, Anal. Chem. 1994, 66, 1424.
    129. Q. Gao, H. M. Pang, E. S. Yeung, Electrophoresis, 1999, 20, 1518.

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