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系統識別號 U0002-1407200813022400
中文論文名稱 微型整合分析系統結合電化學偵測左旋多巴、多巴胺及多巴克
英文論文名稱 Simultaneous determination of L-DOPA, dopamine and DOPAC with electrochemical based micro total analysis system, µ-TAS
校院名稱 淡江大學
系所名稱(中) 化學學系碩士班
系所名稱(英) Department of Chemistry
學年度 96
學期 2
出版年 97
研究生中文姓名 陳信良
研究生英文姓名 Hsin-Liang Chen
學號 693170549
學位類別 碩士
語文別 中文
口試日期 2008-06-03
論文頁數 101頁
口試委員 指導教授-林孟山
委員-蔡東湖
委員-何佳安
委員-施偉仲
中文關鍵字 微晶片  聚二甲基矽氧烷  電化學偵測  安培法 
英文關鍵字 microchip  PDMS  electrochemical detection  amperometry 
學科別分類 學科別自然科學化學
中文摘要 相較於過去以電漿處理、動態塗佈、共價鍵修飾和化學氣相沈積法等修飾微全偵測系統的方法,本研究中發展新型具有快速、可靠又穩定的方法去修飾微全偵測系統以提升純聚二甲基矽氧烷的電滲透流強度。實驗過程中用微全偵測系統去偵測最重要之一的神經傳導物質-多比胺和它的代謝物-多巴克和最常使用在治療巴金森氏症的藥物-左旋多巴。修飾的方法是將鄰苯二甲酸氫鉀和聚二甲基矽氧烷混合、硬化成型即可。對於未修飾過的純聚二甲基矽氧烷微流道而言,在分離三種分析物的解析度只有0.57,但在初步探討中修飾0.10% (w/w)鄰苯二甲酸氫鉀的微流道分離分析物時解析度提升為1.01。在最佳化的條件下做20次連續偵測,訊號的相對標準偏差分別為1.65%、2.10% 和 2.82%。
英文摘要 In contrast to the prior schemes of plasma treatment, dynamic coating, covalent bonding, and chemical vapor deposition (CVD), in preparation of micro total analysis system (μ-TAS), a prompt, reliable, and permanent scheme to enhance the electroosmotic flow of native PDMS was described. The results indicate the relative standard deviations for dopamine, levodopa, and DOPAC are 1.65%, 2.10%, and 2.82%, respectively. Here, we intend to develop a new μ-TAS to determine one of the most important neurotransmitters, dopamine, and its metabolite, DOPAC, and the most frequently used therapeutic medication, levodopa. The modification of electroosmotic flow (EOF) was simply mixing together with 12 milligrams of potassium biphthalate (KHP) and 10 grams of PDMS. In contrast to the native PDMS with resolution (RS) of 0.57, the 0.10%(w/w) KHP modified PDMS microchannel was used to separate dopamine, levodopa, and DOPAC, with improved resolution of 1.01 between dopamine and levodopa. In the mean time, the EOF was also studied by the retention time of neutral molecule, catechol, and the contact angle studies.
論文目次 第一章 序論......................................................................................................... 1
1-1 研究背景....................................................................................................... 1
1-2 微全分析系統製作方法............................................................................... 1
1-2-1 奈米結構掃描式探針法........................................................................ 1
1-2-2 軟式光蝕刻法........................................................................................ 2
1-2-2-1 鑄模複印法........................................................................................ 2
1-2-2-2 微接觸印刷法..................................................................................... 3
1-2-2-3 溶劑輔助微模鑄法............................................................................. 4
1-2-3 熱壓法.................................................................................................... 5
1-2-4 形貌直接光刻法.................................................................................... 6
1-2-5 中性介穩定態原子光刻法.................................................................... 6
1-3 底材的種類與選擇....................................................................................... 6
1-4 底材修飾....................................................................................................... 7
1-4-1 能量曝照改質法.................................................................................... 8
1-4-2 動態溶液修飾法.................................................................................... 9
1-4-3 聚合電解質多層修飾法........................................................................ 10
1-4-4 共價鍵修飾法........................................................................................ 10
1-4-5 鈰催化聚合反應修飾法........................................................................ 11
1-4-6 矽烷化修飾法........................................................................................ 12
1-4-7 化學氣相沈積修飾法............................................................................ 13
1-5 微全分析系統............................................................................................... 13
1-5-1 樣品注入模式........................................................................................ 14
1-5-1-1 夾擠注入法........................................................................................ 15
1-5-1-2 漂浮注入法........................................................................................ 16
1-5-1-3 閘閥注入法........................................................................................ 17
1-5-2 樣品分離的原理.................................................................................... 18
1-5-2-1 電泳流................................................................................................ 19
1-5-2-2 電滲透流............................................................................................ 20
1-5-3 樣品偵測方法........................................................................................ 22
End-channel detection............................................................................. 23
In-channel detection................................................................................ 24
Off-channel detection............................................................................. 25
1-5-4 Decoupler................................................................................................ 26
1-6 神經傳導物質簡介....................................................................................... 28
1-6-1 巴金森氏症............................................................................................ 28
1-6-2 神經傳導物質代謝途徑........................................................................ 29
1-7 儀器控制與資料擷取之圖形化程式語言-LabVIEW................................. 30
1-8 本研究的目的............................................................................................... 31

第二章 實驗部份................................................................................................ 32
2-1儀器與設備.................................................................................................... 32
2-2 藥品............................................................................................................... 33
2-3 網印厚膜碳電極的製備............................................................................... 34
2-4 微流道的製備............................................................................................... 35
2-4-1 光罩之製作........................................................................................... 35
2-4-2 負光阻式陽模之詳細程序製作........................................................... 35
2-4-3 造模法(casting)製作PDMS微流道..................................................... 37
2-4-4 系統裝置............................................................................................... 38
2-5 接觸角及比導電度的量測........................................................................... 39
2-6 電滲透流的量測........................................................................................... 40
2-7 樣品的製備................................................................................................... 41
2-8 微透析實驗................................................................................................... 41
2-9 實驗條件的設計........................................................................................... 42
2-9-1 酸鹼值之探討........................................................................................ 42
2-9-2 緩衝溶液的種類.................................................................................... 43
2-9-3 緩衝溶液的濃度.................................................................................... 43
2-9-4 偵測過電位的探討................................................................................ 44
2-9-5 PDMS摻雜Potassium Biphthalate作修飾的比例之探討.................... 44
2-9-6 晶片電泳參數最佳化之探討................................................................ 44
2-10 分析特性..................................................................................................... 45
2-11透析液的量測………………………………………………………………………………………… 45

第三章 結果與討論................................................................................ 46
3-1 偵測機制...................................................................................................... 46
3-2 偵測條件最佳化探討.................................................................................. 52
3-2-1 酸鹼值之探討....................................................................................... 52
3-2-2 緩衝溶液的種類................................................................................... 56
3-2-3 緩衝溶液的濃度................................................................................... 59
3-2-4 偵測過電位的探討............................................................................... 63
3-2-5 PDMS摻雜Potassium Biphthalate作修飾的比例之探討.................... 66
3-2-6 進樣時間之探討................................................................................... 71
3-2-7 分離電壓之探討................................................................................... 74
3-3 分析特性...................................................................................................... 77
3-4 透析液的量測............................................................................................. 83
3-5 結論.............................................................................................................. 85
參考文獻.............................................................................................................. 93






表目錄
表(一) 初步修飾與純PDMS探討之訊號值、滯留時間、解析度及接觸角.......... 49
表(二) 酸鹼值探討之訊號值、滯留時間及比導電度............................................ 53
表(三) 緩衝溶液種類探討之訊號值、滯留時間、解析度及比導電度.................. 59
表(四) 緩衝溶液濃度探討之訊號值、滯留時間及比導電度................................ 60
表(五) 偵測過電位探討之訊號值及滯留時間...................................................... 66
表(六) PDMS摻雜Potassium biphthalate作修飾的比例探討之訊號值、 滯留時間和解析度..................................................................................... 67
表(七) PDMS摻雜Potassium biphthalate作修飾的比例探討之接觸角 及電滲透流遷移率..................................................................................... 71
表(八) 進樣時間探討之訊號值和滯留時間.......................................................... 74
表(九) 分離電壓探討之訊號值、滯留時間和解析度............................................ 77
表(十) 偵測系統之最佳化條件與分析特性.......................................................... 82

圖目錄
圖1-1 鑄模複印法的流程圖................................................................................... 3
圖1-2 自組裝過程示意圖....................................................................................... 4
圖1-3 使用紫外線修飾PDMS流道表面接枝的聚合反應................................... 11
圖1-4 鈰催化聚合反應........................................................................................... 12
圖1-5 夾擠注入法(a)樣品注入模式(b)樣品分離模式.......................................... 15
圖1-6 飄浮注入法(a)樣品注入模式(b)樣品分離模式.......................................... 16
圖1-7 閘閥注入法(a)樣品注入前模式(b)樣品注入模式(c)樣品分離模式......... 18
圖1-8 管徑表面電雙層示意圖............................................................................... 21
圖1-9 電泳中電滲流推動帶正負電及中性物質的示意圖................................... 22
圖1-10 Levodopa的代謝途徑................................................................................. 30
圖2-1 Aqueous-based polyurethane聚合分子結構................................................ 34
圖2-2 負光阻式陽模製程....................................................................................... 36
圖2-3 造模法製程圖............................................................................................... 37
圖2-4 系統裝置圖................................................................................................... 38
圖2-5 接觸角量測示意圖....................................................................................... 39
圖2-6 微透析管示意圖.......................................................................................... 42
圖3-1 Catecholamine的氧化反應機制................................................................... 47
圖3-2 以網印厚膜碳電極所得之典型循環伏安法............................................... 50
圖3-3 未修飾(A)及修飾Potassium biphthalate(B) PDMS對電流特性影響之電泳圖.............................................................................................................. 51
圖3-4 酸鹼值對dopamine(a)、levodopa(b)和DOPAC(c)訊號值影響之探討........ 54
圖3-5 酸鹼值對dopamine(a)、levodopa(b)和DOPAC(c)訊號值影響之電泳圖.... 55
圖3-6 緩衝溶液種類對dopamine(a)、levodopa(b)和DOPAC(c)訊號值影響之探討.............................................................................................................. 57
圖3-7 緩衝溶液種類對dopamine(a)、levodopa(b)和DOPAC(c)訊號值影響之電泳圖........................................................................................................... 58
圖3-8 緩衝溶液濃度對dopamine(a)、levodopa(b)和DOPAC(c)訊號值影響之探討............................................................................................................... 61
圖3-9 緩衝溶液濃度對dopamine(a)、levodopa(b)和DOPAC(c)訊號值影響之電泳圖........................................................................................................... 62
圖3-10 偵測過電位對dopamine(a)、levodopa(b)和DOPAC(c)訊號值影響之 探討............................................................................................................. 64
圖3-11 偵測過電位對dopamine(a)、levodopa(b)和DOPAC(c)訊號值影響之 電泳圖…………………………………………………………………………………………………… 65
圖3-12 PDMS摻雜Potassium biphthalate作修飾物的比例對dopamine(a)、levodopa(b)和DOPAC(c)訊號值影響之探討.............................................. 68
圖3-13 PDMS摻雜Potassium biphthalate作修飾物的比例對dopamine(a)、levodopa(b)和DOPAC(c)訊號值影響之電泳圖.......................................... 69
圖3-14 PDMS摻雜Potassium biphthalate作修飾物比例對電滲透流影響之 探討............................................................................................................. 70
圖3-15 進樣時間對dopamine(a)、levodopa(b) 和DOPAC(c) 訊號值影響之 探討............................................................................................................. 72
圖3-16 進樣時間對dopamine(a)、levodopa(b) 和DOPAC(c) 訊號值影響之 電泳圖......................................................................................................... 73
圖3-17 分離電壓對dopamine(a)、levodopa(b) 和DOPAC(c) 訊號值影響之 探討............................................................................................................. 75
圖3-18 分離電壓對dopamine(a)、levodopa(b) 和DOPAC(c) 訊號值影響之 電泳圖......................................................................................................... 76
圖3-19 PDMS晶片電泳偵測dopamine(a)、levodopa(b) 和DOPAC(c) 之 校正曲線圖................................................................................................. 79
圖3-20 PDMS晶片電泳偵測dopamine(a)、levodopa(b)和DOPAC(c)之電泳圖..... 80
圖3-21 連續偵測20次之系統穩定性.................................................................... 81
圖3-22在透析液中添加已知濃度dopamine、levodopa及DOPAC之電泳圖........ 84


附 錄
附錄1 負光阻式陽模之製作程序.......................................................................... 86
附錄2電壓控制之操作介面.................................................................................. 87
附錄3電壓控制之程式內容.................................................................................. 88
附錄4 Labview控制高電壓繼電器之電路圖......................................................... 91
附錄5 Dopamine、L-DOPA、DOPAC的解離常數圖.............................................. 92
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