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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
相關次數： 點閱：394  下載：4 
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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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