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研究生:蔡毅軒
研究生(外文):Yi-Hsuan Tsai
論文名稱:利用LVSS-ASEI-Sweeping線上濃縮技術分離四氫大麻酚與其主要代謝物
論文名稱(外文):Separation of △9-tetrahydrocannabinol and its major metabolites by large-volume sample stacking-anion selective exhaustive injection-sweeping
指導教授:林宜慧林宜慧引用關係
學位類別:碩士
校院名稱:中國醫藥大學
系所名稱:藥學系碩士班
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2013
畢業學年度:102
語文別:中文
論文頁數:54
中文關鍵詞:大麻四氫大麻酚毛細管電泳線上濃縮大體積樣品進樣陰離子選擇性電動進樣
外文關鍵詞:capillary electrophoresison-line concentrationcannabistetrahydrocannabinolTHCTHC-OH-THC-COOHASEILVSSSweeping
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大麻由植物Cannabis Sativa的葉所製成,由於容易取得及較低成癮性的關係,為全世界最常被濫用的藥物之一。四氫大麻酚(△9-tetrahydrocannabinol, THC)為大麻中的主要具有神經活性的成分,代謝後於人體含量最豐富的代謝物為 11-hydroxy-△9-tetrahydrocannabinol (THC-OH)和11-nor-9-carboxy-△9-tetrahydrocannabinol (THC-COOH)。在此,我們建立了一個線上濃縮分析方法-大體積樣品堆積搭配陰離子選擇性電動進樣掃集法(LVSS-ASEI-Sweeping)針對THC及其代謝物進行分析研究,首先將內含40 %甲醇及100 mM SDS磷酸鹽緩衝溶液(30 mM, pH 2.5)注入毛細管中,接著注入一段高導電度磷酸鹽緩衝溶液(200 mM, 0.5 psi, 1 min, pH 2.5),再以壓力0.5 psi將壓力進樣樣品溶液注入3 min,之後以逆向電壓2.5 kV對待測物取樣6 min,最後利用逆向電壓15 kV進行掃集及分離,於214 nm波長下偵測。此分析方法進行確效後,THC與THC-OH的定量範圍為40~6000 ng/mL,而THC-COOH為2~300 ng/mL,其相關係數皆大於0.9964。各偵測極限為THC 10 ng/mL,THC-OH 5 ng/mL,THC-COOH 500 pg/mL,以THC-COOH比較本法與micellar-electrokinetic chromatography (MEKC)法,本法偵測靈敏度約可提高2000倍。本分析方法優化後可用於尿液檢品分析。本法對大麻檢測提供簡單、快速、高靈敏的分析方法。

Marijuana is made by the leaves of Cannabis Sativa. With its accessible and less addictive, it is the most commonly abused drug in the world. △9-tetrahydrocannabinol (THC) is the major psychoactive component in marijuana. 11-hydroxy-△9-tetrahydrocannabinol (THC-OH) and 11-nor-9-carboxy-△9-tetrahydrocannabinol (THC-COOH) are most abundant metabolites in human after metabolism. Here, we describe a large-volume sample stacking-anion selective exhaustive injection sweeping method for the determination of THC and its metabolites. First, phosphate buffer (30 mM, pH 2.5) containing 40 % methanol and 100 mM SDS are filled into an uncoated fused silica capillary, and then high conductivity phosphate buffer (200 mM, 0.5 psi, 1 min, pH 2.5) was followed. Samples were loaded by pressure injection (0.5 psi, 3 min) before electrokinetic injection (-2.5 kV, 6 min). The stacking step and separation were performed at -15 kV. The detection wavelength was set at 214 nm. During method validation, calibration polts were linear (r≧0.9964) over a range of 40~6000 ng/mL for THC and THC-OH, and 2~300 ng/mL for THC-COOH. The limits of detection (LODs, S/N=3) were 10 ng/mL for THC, 5 ng/mL for THC-OH, and 500 pg/mL for THC-COOH. Comparing with micellar-electrokinetic chromatography, the results indicated that this stacking CE method could increase about 2000-fold sensitivity for the analysis of THC-COOH. After optimization, this method was feasible to detect THC and it’smetabolites in human urine. It can provide advantages such as simplicity, rapidity, and high sensitivity for the analysis of THC.

第一章 緒論 1
第二章 實驗方法 9
第一節 試藥及材料 9
第二節 使用之主要儀器 10
第三節 試藥溶液配製 11
1. 儲備溶液 11
2. 分析作業用溶液之配製 11
3. 緩衝溶液的配製 12
4. 檢量線之作業溶液 13
5. 同日間與異日間精確度及準確性試驗之作業溶液 14
6. 尿液之前處理 15
7. 尿液之萃取 15
第四節 毛細管清洗方式 16
第三章 結果及討論 17
第一節 基本分析條件之建立 17
1. 緩衝溶液之濃度 17
2. 緩衝溶液之pH值 19
3. 有機溶媒修飾劑的種類及比例 21
4. HCB濃度之探討 24
5. 緩衝溶液中SDS濃度之探討 26
6. 大體積樣品進樣的時間 28
7. 電動取樣的電壓 30
8. 電動取樣的時間 32
9. 電動取樣用樣品溶液之溶媒種類 34
10. 電動取樣用樣品溶液中的ACN比例 36
11. 分離電壓 38
12. 待測物THC、THC-OH、THC-COOH之定位 40
第二節 毛細管電泳條件 42
第三節 靈敏度提升效果之比較 43
第四節 基本分析方法之確效 44
1. THC與其代謝物之定量 44
2. 分析方法的精密度及準確性之探討 46
第五節 應用分析 48
第六節 結論 50
第四章 參考文獻 51
附錄 人體試驗/研究同意證明書


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