臺灣博碩士論文加值系統

安非他命類藥物為一種中樞神經刺激藥物；愷他命則為近年來新興之濫用藥物，此二種藥物常被青少年混合吸食，同時；分析濫用藥物之鏡像異構物是相當重要的ㄧ個課題。本研究以毛細管電泳搭配光電二極體陣列偵測器，在三種不同的基質(甲醇/水溶液、尿液及血液)進行安非他命類及愷他命藥物的非鏡像及鏡像異構物的分析。其分析藥物包含安非他命、甲基安非他命、亞甲基雙氧安非他命、亞甲基雙氧甲基安非他命、亞甲基雙氧乙基安非他命、愷他命及去甲基愷他命。藥物分析使用的電泳緩衝溶液為tris-(100 mM)，pH 3.5以及未塗覆物質而長度為50.2 cm和內徑為50 �慆的毛細管；待測藥物的分析滯留時間皆在15分鐘內。並且，線性回歸數值均高於0.996，其添加回收率落在100±10％的可信範圍內。
進行鏡像異構物分析時，在電泳緩衝溶液中添加β–環糊精作為鏡像異構物選擇劑，其餘分析條件則與非鏡像異構物的分析條件相同。待測藥物的鏡像異構物之分析滯留時間皆在20分鐘內。並且，線性回歸數值均高於0.9960，其回收率亦在100±10％的可信範圍內。此外，在三種基質中無論是在鏡像或非鏡像之結構分析上，其精密度 (％RSD)及準確度 (％Bias)上的驗證皆小於10％的可接受範圍。
以毛細管電泳分析儀，偵測濫用藥物及其鏡像異構物，不但快速簡便而且解析度高，可彌補氣相層析質譜儀在分析時必須事先作複雜的衍生工作才能達成目的之不足。期望此方法可實際應用於法庭和臨床檢驗上的分析。

Amphetamines are kinds of medicine that stimulates central nervous system, whereby ketamine has been assigned emerging abuses in the recent years. These two kinds of medicine were frequently mixed and abused by young ages. The analysis of chiral for the analyze is another critical issue. In this study, a capillary electrophoresis (CE) method with photodiode array detector (DAD) for the detection of amphetamines and ketamines in methanol/water solution, urine, and blood has been investigated including amphetamine (A), methamphetamine (MA), methylenedioxy-amphetamine (MDA), methylenedioxy-methamphetamine (MDMA), methylenedioxy-ethylamphetamine (MDEA), ketamine, and norketamine. Achiral analysis were separated within 15 minutes through an uncoated fused-silica capillary (50.2cm x 50µm) using tris-phosphate buffer (pH=3.5, 100 mM). Correlation coefficient was higher than 0.9960 in each sample. The recoveries in this method were determined to be 100±10%.
Beside the addition of β-cyclodextrin (20mM, as a chiral selector) in tirs-phosphate buffer for the separation of chiral analyze, the condition of CE for chiral analyze are the same as that for achiral analyze. Correlation coefficient was remained to be higher than 0.9960 in each samples. The recoveries in this method were determined to be 100±10%. Precision and accuracy for the analytic method were smaller than 10% in three matrixes.
Analysis of chiral and achiral isomers of illegal drugs with capillary electrophoresis takes the advantages to high resolution and rapid dectection. Furthermore, the method is complementary to GC/MS for analysis of amphetamines and ketamines in biological samples. Eventually, the results obtained in this study contribute to the analysis of drug abused in forensic sciences and clinical laboratories.

中文摘要 ------------------------------------------------------------------ i

英文摘要 ------------------------------------------------------------------ iii

目錄 ------------------------------------------------------------------ v

表目錄 ------------------------------------------------------------------ viii

圖目錄 ------------------------------------------------------------------ x

縮寫表 ------------------------------------------------------------------ xii

第1章 緒論------------------------------------------------------------ 1
第2章 文獻探討------------------------------------------------------ 3
2.1 安非他命及其相關衍生物之探討------------------------ 3
2.1.1 安非他命(Amphetamine)----------------------------------- 3
2.1.2 甲基安非他命(Methamphetamine)----------------------- 3
2.1.3 雙氧甲基安非他命(Methylene-dioxyamphetamine)--------------------------
4
2.1.4 亞甲二氧基甲基安非他命(Methylene-dioxymethamphetamine)---------------------
5
2.1.5 亞甲二氧基乙基安非他命(Methylene-dioxyethylamphetamine)--------------------
5
2.1.6 安非他命類藥物對健康之危害--------------------------- 6
2.2 愷他命之探討------------------------------------------------ 7
2.2.1 愷他命(ketamine)------------------------------------------- 7
2.2.2 愷他命對健康之危害--------------------------------------- 7
2.3 鏡像異構物的分析探討------------------------------------ 8
2.3.1 背景介紹----------------------------------------------------- 8
2.3.2 鏡像異構物分離模式--------------------------------------- 9
2.3.3 環糊精 (鏡像選擇劑)-------------------------------------- 9
2.4 濫用藥物分析方法------------------------------------------ 11
2.5 毛細管電泳分析簡介--------------------------------------- 15
2.6 研究動機------------------------------------------------------ 19
第3章 材料與方法--------------------------------------------------- 21
3.1 實驗材料------------------------------------------------------ 21
3.1.1 藥物標準品及內部標準品--------------------------------- 21
3.1.2 實驗藥品------------------------------------------------------ 21
3.1.3 其他實驗器材------------------------------------------------ 22
3.1.4 實驗儀器------------------------------------------------------ 22
3.2 實驗方法------------------------------------------------------ 23
3.2.1 溶液配製------------------------------------------------------ 23
3.2.2 檢體前處理--------------------------------------------------- 23
3.2.3 分析條件與步驟--------------------------------------------- 25
3.2.4 鏡像異構物分析條件之差異性--------------------------- 25
3.3 方法建立及驗證--------------------------------------------- 26
3.3.1 檢量線建立--------------------------------------------------- 26
3.3.2 準確度--------------------------------------------------------- 26
3.3.3 精密度--------------------------------------------------------- 27
3.3.4 添加回收率--------------------------------------------------- 27
3.3.5 偵測極限------------------------------------------------------ 28
3.3.6 臨床檢體之分析--------------------------------------------- 28
第4章 研究結果------------------------------------------------------ 29
4.1 非鏡像異構物之分析--------------------------------------- 29
4.2 鏡像異構物之分析------------------------------------------ 32
4.3 臨床檢體之分析--------------------------------------------- 37
第5章 討論與結論--------------------------------------------------- 38
5.1 分析方法確立------------------------------------------------ 38
5.2 分析結果與相關文獻比較--------------------------------- 39
第6章 參考文獻------------------------------------------------------ 43
附表 ------------------------------------------------------------------ 51
附圖 ------------------------------------------------------------------ 68

1.Amini, A.; Sorman, U. P.; Lindgren, B. H., and Westerlund, D. Enantioseparation of anaesthetic drugs by capillary zone electrophoresis using cyclodextrin-containing background electrolytes. Electrophoresis. 1998; 19(5): 731–737.
2.Anastos, N.; Barnett, N.W. and Lewis, S.W. Capillary electrophoresis for forensic drug analysis: A review. Talanta. 2005; 67(3): 269–279.
3.Armbruster, D. A.; Tillman, M.D., and Hubbs, L.M. Limit of detection (LQD)/limit of quantitation (LOQ): comparison of the empirical and the statistical methods exemplified with GC-MS assays of abused drugs. Clin Chem. 1994; 40(7 Pt 1): 1233–1238.
4.Blanco, M. and Valverde, I. Choice of chiral selector for enantioseparation by capillary electrophoresis. Trends Analyt Chem. 2003; 22(7–8): 428–439.
5.Boatto, G.; Faedda, M. V.; Pau, A.; Asproni, B.; Menconi, S., and Cerri, R. Determination of amphetamines in human whole blood by capillary electrophoresis with photodiode array detection. J Pharm Biomed Anal. 2002; 29(6): 1073–1080.
6.Bressolle, F.; Bromet-Petit, M., and Audran, M. Validation of liquid chromatographic and gas chromatographic methods. Applications to pharmacokinetics. J Chromatogr B Biomed Appl. 1996; 686(1): 3–10.
7.Caplan, Y. H. and Levine, B. Abbott phencyclidine and barbiturates abused drug assays: evaluation andcomparison of ADx FPIA, TDx FPIA, EMIT, and GC/MS methods. J Anal Toxicol. 1989; 13(5): 289–292.
8.Chankvetadze, B. Enantioseparations by using capillary electrophoretic techniques. The story of 20 and a few more years. J Chromatogr A. 2007; 1168(1–2): 45–70.
9.Cherkaoui, S. and Veuthey, J. L. Use of negatively charged cyclodextrins for the simultaneous enantioseparation of selected anesthetic drugs by capillary electrophoresis-mass spectrometry. J Pharm Biomed Anal. 2002; 27(3–4): 615–626.
10.Choi, J. G.; Kim, M.; Dadoo, R., and Zare, R. N. Electrophoretron: a new method for enhancing resolution in electrokinetic separations. J Chromatogr A. 2001; 924(1–2): 53–58.
11.Chung, Y. L.; Liu, J. T., and Lin, C. H. On-line identification of 3,4-methylenedioxymethamphetamine in human urine by non-aqueous capillary electrophoresis-fluorescence spectroscopy at 77 K. J Chromatogr B Biomed Sci Appl. 2001; 759(2): 219–226.
12.Colbert, D. L. and Childerstone, M. Multiple drugs of abuse in urine detected with a single reagent and fluorescence polarization. Clin Chem. 1987; 33(10): 1921–1923.
13.Colbert, D. L.; Gallacher, G., and Mainwaring-Burton, R. W. Single-reagent polarization fluoroimmunoassay for amphetamine in urine. Clin Chem. 1985; 31(7): 1193–1195.
14.Colbert, D. L.; Smith, D. S.; Landon, J., and Sidki, A. M. Single-reagent polarization fluoroimmunoassay for the cocaine metabolite, benzoylecgonine, in urine. Ann Clin Biochem. 1986; 23 ( Pt 1): 37–41.
15.Colbert, D. L.; Smith, D. S.; Landon, J. and Sidki, A. M. Single-reagent polarization fluoroimmunoassay for barbiturates in urine. Clin Chem. 1984; 30(11): 1765–1769.
16.Del Mar Ramirez Fernandez, M.; Laloup, M.; Wood, M.; De Boeck, G.; Lopez-Rivadulla, M.; Wallemacq, P., and Samyn, N. Liquid chromatography-tandem mass spectrometry method for the simultaneous analysis of multiple hallucinogens, chlorpheniramine, ketamine, ritalinic acid, and metabolites, in urine. J Anal Toxicol. 2007; 31(8): 497–504.
17.Desiderio, C.; Ossicini, L., and Fanali, S. Analysis of hydroquinone and some of its ethers by using capillary electrochromatography. J Chromatogr A. 2000; 887(1–2): 489–496.
18.Di Pietra, A. M.; Gotti, R.; Del Borrello, E.; Pomponio, R., and Cavrini, V. Analysis of amphetamine and congeners in illicit samples by liquid chromatography and capillary electrophoresis. J Anal Toxicol. 2001; 25(2): 99–105.
19.Feng, N.; Vollenweider, F. X.; Minder, E. I.; Rentsch, K.; Grampp, T., and Vonderschmitt, D. J. Development of a gas chromatography-mass spectrometry method for determination of ketamine in plasma and its application to human samples. Ther Drug Monit. 1995; 17(1): 95–100.
20.Hudson, J.C.; Golin, M.; Malcolm, M. and Whiting, C.F. Capillary zone electrophoresis in a comprehensive screen for drugs of forensic interest in whole blood: an update, Can Soc Forensic Sci.1998; 31 (1): 1–29
21.Iio, R.; Chinaka, S.; Takayama, N., and Hayakawa, K. Simultaneous chiral analysis of methamphetamine and related compounds by capillary electrophoresis/mass spectrometry using anionic cyclodextrin. Anal Sci. 2005; 21(1): 15–19.
22.Iwata, Y. T.; Inoue, H.; Kuwayama, K.; Kanamori, T.; Tsujikawa, K.; Miyaguchi, H., and Kishi, T. Forensic application of chiral separation of amphetamine-type stimulants to impurity analysis of seized methamphetamine by capillary electrophoresis. Forensic Sci Int. 2006; 161(2–3): 92–96.
23.Koppenhoefer, B.; Epperlein, U.; Christain, B.; Yibing, J.; Yuying, C. and Bingcheng, L. Separation of enantiomers of drugs by capillary electrophoresis I. gama-cyclodextrin as chiral solvating agent. J Chromatogr A. 1995; 717: 191–190.
24.Koppenhoefer, B.; Epperlein, U.; Christian, B.; Lin, B.; Ji, Y., and Chen, Y. Separation of enantiomers of drugs by capillary electrophoresis. III. Beta-cyclodextrin as chiral solvating agent. J Chromatogr A. 1996; 735(1–2): 333–343.
25.Kraemer, T. and Maurer, H. H. Determination of amphetamine, methamphetamine and amphetamine-derived designer drugs or medicaments in blood and urine. J Chromatogr B Biomed Sci Appl. 1998; 713(1): 163–187.
26.Kunsman, G. W.; Levine, B.; Kuhlman, J. J.; Jones, R. L.; Hughes, R. O.; Fujiyama, C. I., and Smith, M. L. MDA-MDMA concentrations in urine specimens. J Anal Toxicol. 1996; 20(7): 517–521.
27.Kunsman, G. W.; Manno, J. E.; Cockerham, K. R., and Manno, B. R. Application of the Syva EMIT and Abbott TDx amphetamine immunoassays to the detection of 3,4-methylene-dioxymethamphetamine (MDMA) and 3,4-methylene-dioxyethamphetamine (MDEA) in urine. J Anal Toxicol. 1990; 14(3): 149–153.
28.Kuroda, N.; Nomura, R.; al-Dirbashi, O.; Akiyama, S., and Nakashima, K. Determination of methamphetamine and related compounds by capillary electrophoresis with UV and laser-induced fluorescence detection. J Chromatogr A. 1998; 798(1–2): 325–334.
29.Larenza, M. P.; Knobloch, M.; Landoni, M. F.; Levionnois, O. L.; Kronen, P. W.; Theurillat, R.; Schatzmann, U., and Thormann, W. Stereoselective pharmacokinetics of ketamine and norketamine after racemic ketamine or S-ketamine administration in Shetland ponies sedated with xylazine. Vet J. 2008; 177: 432–435.
30.Lin, B.; Wuerthner, S.; Epperlein, U. and Koppenhoefer, B. Separation of enantiomers of drugs by capillary electrophoresis Part 8. beta-cyclodextrin as chiral solvating agent. Talanta. 1997; 46: 743–749.
31.Lin, C. E.; Cheng, H. T.; Fang, I. J.; Liu, Y. C.; Kuo, C. M.; Lin, W. Y., and Lin, C. H. Strategies for enantioseparations of catecholamines and structurally related compounds by capillary zone electrophoresis using sulfated beta-cyclodextrins as chiral selectors. Electrophoresis. 2006; 27(17): 3443–3451.
32.Lin, Y. H.; Li, J. H.; Ko, W. K., and Wu, S. M. Direct and sensitive analysis of methamphetamine, ketamine, morphine and codeine in human urine by cation-selective exhaustive injection and sweeping micellar electrokinetic chromatography. J Chromatogr A. 2006; 1130(2): 281–286.
33.Logan, B.K. and Couper, F.J. 3, 4-Methylenedioxymethamphetamine effects on human performance and behavior. Forensic Sci Rev. 2003; 15(1): 12–28.
34.Long, G. L. and Dorn, D.S. Sociologists'' attitudes toward ethical issues: the management of an impression. Sociol Soc Res. 1983;67(3): 288–300.
35.Lu, W. and Cole, R. B. Determination of chiral pharmaceutical compounds, terbutaline, ketamine and propranolol, by on-line capillary electrophoresis-electrospray ionization mass spectrometry. J Chromatogr B Biomed Sci Appl. 1998; 714(1): 69–75.
36.Paterson, S.; McLachlan-Troup, N.; Cordero, R.; Dohnal, M., and Carman, S. Qualitative screening for drugs of abuse in hair using GC-MS. J Anal Toxicol. 2001; 25(3): 203–208.
37.Pizarro, N.; Ortuno, J.; Farre, M.; Hernandez-Lopez, C.; Pujadas, M.; Llebaria, A.; Joglar, J.; Roset, P. N.; Mas, M.; Segura, J.; Cami, J., and de la Torre, R. Determination of MDMA and its metabolites in blood and urine by gas chromatography-mass spectrometry and analysis of enantiomers by capillary electrophoresis. J Anal Toxicol. 2002; 26(3): 157–165.
38.Ramseier, A.; Caslavska, J., and Thormann, W. Stereoselective screening for and confirmation of urinary enantiomers of amphetamine, methamphetamine, designer drugs, methadone and selected metabolites by capillary electrophoresis. Electrophoresis. 1999; 20(13): 2726–2738.
39.Rofael, H. Z. and Abdel-Rahman, M. S. Development and validation of a high-performance liquidchromatography method for the determination of cocaine, its metabolites and ketamine. J Appl Toxicol. 2002; 22(2): 123–128.
40.Tagliaro, F.; Turrina, S.; Pisi, P.; Smith, F. P., and Marigo, M. Determination of illicit and/or abused drugs and compounds of forensic interest in biosamples by capillary electrophoretic electrokinetic methods. J Chromatogr B Biomed Sci Appl. 1998; 713(1): 27–49.
41.Theurillat, R.; Knobloch, M.; Levionnois, O.; Larenza, P.; Mevissen, M., and Thormann, W. Characterization of the stereoselective biotransformation of ketamine to norketamine via determination of their enantiomers in equine plasma by capillary electrophoresis. Electrophoresis. 2005; 26(20): 3942–3951.
42.Theurillat, R.; Knobloch, M.; Levionnois, O.; Larenza, P.; Mevissen, M. and Thormann, W. Characterization of the stereoselective biotransformation of ketamine to norketamine via determination of their enantiomers in equine plasma by capillary electrophoresis. Electrophoresis. 2005; 26(20): 3942–3951.
43.Tsukamoto, T.; Ushio, T., and Haginaka, J. Chiral resolution of basic drugs by capillary electrophoresis with new glycosaminoglycans. J Chromatogr A. 1999; 864(1): 163–171.
44.Wallace, J. E.; Hamilton, H. E.; Schwertner, H.; King, D. E.; McNay, J. L., and Blum, K. Thin-layer chromatographic analysis of cocaine and benzoylecgonine in urine. J Chromatogr. 1975; 114(2): 433–441.
45.Wallenborg, S. R.; Lurie, I. S.; Arnold, D. W., and Bailey, C. G. On-chip chiral and achiral separation of amphetamine and related compounds labeled with 4-fluoro-7-nitrobenzofurazane. Electrophoresis. 2000; 21(15): 3257–3263.
46.Wan, H. and Blomberg, L. G. Chiral separation of amino acids and peptides by capillary electrophoresis. J Chromatogr A. 2000; 875(1–2): 43–88.
47.Wang, F. and Khaledi, M. G. Enantiomeric separations by nonaqueous capillary electrophoresis. J Chromatogr A. 2000; 875(1–2): 277–293.
48.Wu, W. S. and Tsai, J. L. Investigation of the stability of drugs of abuse in urine sample by capillary zone electrophoresis. Chin Pharm J. 1997; 49(4): 239–248.
49.Wang, H.; Gu, J.L.; Hu, H.F.; Dai, R.J.; Ding, T.H. and Fu, R.N. Study on the chiral separation of basic drugs by capillary zone electrophoresis using β-CD and derivatized β-CDs as chiral selectors. Anal Chim Acta. 1998; 359: 39–46.
50.謝茜如，魏國佐，毛細管電動層析之對掌性靜相的發展與應用，中國化學，2001; 59 (3): 409–422。
51.Clarke, E. Clarke''s analysis of drugs and poisons: in pharmaceuticals, body fluids and postmortem material. 2004.
52.Douglas, A.; 方嘉德譯，儀器分析 (精選本)，2000。
53.蔡裕庚，尿液毒品測試簡介，1995。
54.李志�琤D編/衛生署管制藥品管理局，藥物濫用，2002。
55.劉正雄，應用生物藥劑學與藥物動力學，1995。
56.管制藥品管理局線上資料庫，http://www.nbcd.gov.tw/home/dep/subpage7.html
57.管制藥品管理局線上資料庫，http://www.nbcd.gov.tw/home/dep/subpage12.html
58.Psychoactive chemicals, 2007 http://www.erowid.org/chemicals/meth/meth.shtml
59.Psychoactive chemicals, 2007 http://www.erowid.org/chemicals/mdma/mdma.shtml