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研究生: 黃宣銘
Hsuan-Ming Huang
論文名稱: 毛細管電泳/紫光LED誘導螢光偵測法
Determination of Dopamine in Urine by Capillary Electrophoresis-Violet Light Emitting Diode-
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: 2004
畢業學年度: 92
語文別: 中文
論文頁數: 113
中文關鍵詞: 多巴胺紫光發光二極體
英文關鍵詞: dopamine, violet light emitting diode
論文種類: 學術論文
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  • 本研究以紫光/發光二極體(波長410 nm, 2 mW)為螢光激發光源,配合毛細管電泳-線上掃集濃縮技術/誘導螢光偵測法,成功偵測到尿液中微量的多巴胺。這方法是藉由NDA (naphthalene-2,3-dicarboxaldehyde)做為螢光標識試劑,與多巴胺進行衍生反應以後,以螢光偵測的方式進行測量。研究結果發現,以SDS(十二烷基磺酸鈉)為界面活性劑的毛細管微胞電動層析(MEKC)與掃集-毛細管電泳法(sweeping-MEKC)進行分離時,電泳結果所得到的譜峰其理論板數分別為1 × 105、2 × 105;偵測極限為9.1 × 10-7 M與1.2 × 10-8 M。若以sweeping-MEKC的分離模式,且在樣品基質(sample matrix)與背景緩衝溶液(background electrolyte)之間,添加一截甲醇區段,不僅可以延長樣品進樣的體積,使偵測極限到達4.7 × 10-9 M,SDS微胞被甲醇層先破壞後重組的過程之後,理論板數甚至能增加到 1 × 106。

    Uses and limitations of a methanol plug assisted sweeping-micellar electrokinetic chromatography (sweeping-MEKC) method are described. Dopamine labeled-NDA (naphthalene-2,3-dicarboxaldehyde) was selected as the model compound; this new method is also applied to the determination of dopamine in actual urine samples. An inexpensive violet light-emitting-diode (LED) was used for the light source because this is suit for fluorescence excitation. The number of theoretical plates of the analyte was determined to be ~ 1 × 105 and ~ 2 × 105 by means of a MEKC and sweeping-MEKC and this was improved to ~ 1 × 106 when the methanol plug assisted mode was applied. In addition, the detection limit of NDA-labeled dopamine was determined to be 9.1 × 10-7 M and 1.2 × 10-8 M by means of MEKC and sweeping-MEKC and this was improved to 4.7 × 10-9 M when the methanol plug assisted sweeping-MEKC mode was applied.

    中文摘要 Ⅰ 英文摘要 Ⅱ 目錄 Ⅲ、Ⅳ 圖目錄 Ⅴ、Ⅵ 表目錄 Ⅶ 第一章、緒論 1 1-1 研究目的 1 1-2 分析物簡介 4 1-3 發光二極體簡介 6 第二章、分析方法與原理 10 2-1 毛細管電泳分析之發展歷程 10 2-2 毛細管電泳法之基本原理 14 2-2-1 電泳分離與遷移率 14 2-2-2 電滲流(EOF) 16 2-2-3 管柱分離效率 21 2-3 毛細管電泳層析法的分離模式 23 2-3-1 毛細管區帶電泳(CZE) 24 2-3-2 微胞電動層析法(MEKC) 27 2-4 毛細管線上濃縮技術 31 2-4-1 毛細管電泳線上堆積法(stacking) 34 2-4-2 毛細管電泳線上掃集法(sweeping) 36 2-4-3 陽離子選擇完全注射法(CSEI) 38 2-5 固相萃取法 40 第三章、儀器、藥品與實驗方法 42 3-1 自組式毛細管電泳-紫光LED誘導螢光分析儀 42 3-2 自組式螢光光譜儀 45 3-3 儀器及週邊設備列表 47 3-4 使用藥品列表 50 3-5 多巴胺衍生物製備 52 3-6 尿液前處理 53 3-6-1 尿液的水解 53 3-6-2 尿液的萃取:固相萃取 53 3-7 偵測極限之計算 57 第四章、毛細管電泳/紫光LED誘導螢光法對Dopamine之研究 58 4-1 Dopamine衍生反應之探討 58 4-2 Dopamine衍生物光譜性質之測量 61 4-3 紫光LED發光光譜圖之測量 63 4-4 Dopamine衍生物與紫光LED相關光譜之比較 64 4-5 微胞電動層析法(MEKC)之電泳條件探討 65 4-5-1 最佳化MEKC電泳條件確立 65 4-5-2 MEKC檢量線製作 70 4-6 線上濃縮技術應用:Sweeping-MEKC 72 4-6-1 最佳化Sweeping-MEKC之電泳條件確立 72 4-6-2 Sweeping-MEKC檢量線製作 81 4-6-3甲醇栓(Methanol plug)對Sweeping-MEKC的影響 83 4-6-4 Methanol plug assisted Sweeping-MEKC檢量線製作 91 4-6-5 MEKC、Sweeping-MEKC與Methanol plug assisted sweeping-MEKC 93 4-7 尿液中dopamine的測定 99 4-8-1 最佳電泳條件測試 99 4-8-2尿液中dopamine的偵測 103 第五章、結論 105 參考文獻 107 發表論文 113

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