本研究主要於發展微晶片電泳與電化學偵測之結合，在微晶片的製作過程中，首先利用微積電製程技術於矽晶圓的表面製造出微流道的陽模，再使用poly(dimethylsiloxane) (PDMS)採用造模法的方式翻模製造出微流道，本法的優點有材料成本較低、製作過程簡便適合大量製造等。就電化學偵測而言，本系統利用網版印刷碳電極作為工作電極，並將此工作電極垂直置於微流道之末端以進行偵測分析物，具有可簡單且快速的更換工作電極之優點。
此系統透過於PDMS的製作過程中混入的cellulose acetate以改善分離acetaminophen與p-aminophenol的拖尾現象，而系統在最佳化的條件下，溶液組成20mM 2-(4-Morpholino)ethanesulfonic acid (MES)緩衝溶液pH5.5，內含1mM 氯化鈉，以偵測電位900mV(vs. Ag/AgCl)偵測自電泳分離的acetaminophen與p-aminophenol，偵測時工作電極與流道末端距離為60μm，分離的條件為樣品注入時間3秒，分離電壓2000V，分離時樣品槽與樣品廢液槽所施加的電壓控制在分離電壓80%的水準，所得之分析特性分別為：p-aminophenol的線性範圍從5μM到1.25mM(R=0.999)，偵測靈敏度為0.32nA/μM，acetaminophen的線性範圍從10μM到1mM(R=0.995)，偵測靈敏度為0.17nA/μM，重複17次實驗操作對200μM p-aminophenol與acetaminophen水溶液的偵測，所得之相對標準偏差分別為3.22%與3.62%。

This research demonstrates the feasibility of using microchip for acetaminophen and p-aminophenol with thick film printed electrochemical detector. First, we used the microelectromechanical systems (MEMS) technology to create a positive pattern on the silicon wafer, and then the poly(dimethylsiloxane) (PDMS) microchannel was formed by casting. The PDMS owns many advantages such as low cost, readily mechanical processing and mass replication. For electrochemical detector, the working electrode was fabricated by screen printed technology and subsequently the electrode was mounted perpendicularly to the outlet of microchannel. The advantage of the electrode is easy and 
fast replacement.
The peak spreading of acetaminophen and p-aminophenol was improved by blending 0.2% cellulose acetate for the PDMS. Acetaminophen and p-aminophenol was measured by using amperometry at the optimum condition at buffer solution: 20mM 2-(4-Morpholino)ethanesulfonic acid (MES) buffer pH5.5 containing 1mM sodium chloride; detection potential: 900mV (vs. Ag/AgCl); the distance between the working electrode and microchannel outlet : 60μm; injection time: 3s; the hold voltage in sample and sample waste reservoir during separation: 80% of separation voltage; separation voltage: 2000V. According to optimum operation conditions, the linear ranges of acetaminophen and p-aminophenol are obtained between 10μM to 1mM(R=0.995) and 5μM to 1.25mM(R=0.999), respectively. The sensitivities are 0.17nA/μM for acetaminophen and 0.32nA/μM for p-aminophenol. The detection limits of acetaminophen and p-aminophenol are 3.5μM and 1.8μM(S/N=3), respectively. The relative standard deviation of seventeen repetitive detections are 3.62% for 
acetaminophen and 3.22 for p-aminophenol.

第一章 序論..............................................................................................1
1-1研究背景..............................................................................................1
1-2 毛細管電泳的原理.............................................................................2
1-2-1 電泳流.........................................................................................2
1-2-2 電滲透流.....................................................................................3
1-3 微晶片電泳的發展.............................................................................6
1-3-1 微流道種類與製作方式.............................................................7
1-3-1-1熱壓法........................................................................................9
1-3-1-2雷射燒除法……......................................................................11
1-3-1-3灌模法......................................................................................12
1-3-1-4造模法......................................................................................14
1-4 樣品注入之方式...............................................................................16
1-4-1漂浮注入法.................................................................................16
1-4-2閘閥注入法.................................................................................18
1-4-3 夾擠注入法...............................................................................19
1-5 微晶片之偵測方式...................................................................20
1-5-1 UV/Visible..................................................................................21
1-5-2螢光.............................................................................................22
1-5-3電化學法.....................................................................................23
1-6 acetaminophen與p-aminophenol的重要性......................................30
1-7研究目的........................................…................................................31

第二章 實驗部分....................................................................................33
2-1 儀器與設備.......................................................................................33
2-2藥品....................................................................................................34
2-3網印電極的製備................................................................................35
2-4微流道的製備....................................................................................35
2-4-1光罩之製作.................................................................................35
2-4-2負光阻陽模之製作.....................................................................35
2-4-3造模法(casting)製造PDMS微流道...........................................36
2-4-4系統裝置.....................................................................................37
2-5樣品的製備........................................................................................37
2-6實驗條件的設計................................................................................38
2-6-1緩衝溶液pH............................................................................38
2-6-2緩衝溶液的種類.....................................................................38
2-6-3緩衝溶液的濃度.....................................................................39
2-6-4偵測電位.................................................................................39
2-6-5微流道與偵測電極之距離.....................................................39
    2-6-6毛細管電泳參數最佳化.........................................................39
2.7分析特性.............................................................................................40

第三章 結果與討論................................................................................41
3-1偵測系統之架設...............................................................................41
3-2偵測機制............................................................................................48
3-3 偵測條件最佳化探討.......................................................................52
3-3-1緩衝溶液pH的探討...............................................................52
3-3-2緩衝溶液種類的探討.............................................................56
3-3-3緩衝溶液濃度的探討.............................................................59
3-3-4偵測電位的探討.....................................................................62
3-3-5微流道與偵測電極之距離的探討.........................................64
    3-3-6進樣時間的探討.....................................................................67
3-3-7分離電壓的探討.....................................................................68
3-4 分析特性...........................................................................................77
3-5 結論...................................................................................................82
參考資料..................................................................................................92







圖表目錄
圖3-1：以網印電極所得之典型循環伏安圖..........................................42
圖3-2：以計時電流法偵測10mM多巴胺之晶片電泳圖.......................43
圖3-3：以計時電流法偵測blank(a)、0.1(b)和0.3mM(c)多巴胺之晶片電泳圖......................................................................................................46
圖3-4：工作電極與流道末端距離對氧化電流訊號之影響..................46
圖3-5：固定spacer位置對氧化電流訊號之影響。.................................47
圖3-6：以網印電極所得之典型循環伏安圖..........................................49
圖3-7：(a)未修飾及(b)修飾0.2% cellulose acetate之PDMS對滯留時間影響之實際電流訊號..........................................................................51
圖3-8：溶液酸鹼值之探討......................................................................54
圖3-9：溶液酸鹼值改變下之實際電流響應訊號..................................55
圖3-10：緩衝溶液種類探討....................................................................57
圖3-11：緩衝溶液類別探討之實際電流響應訊號................................58
圖3-12：緩衝溶液濃度的探討................................................................60
圖3-13：MES緩衝溶液於不同溶液濃度下之實際電流響應訊號........61
圖3-14：偵測電位的探討........................................................................63
圖3-15：微流道末端與偵測電極之距離探討........................................65
圖3-16：微流道末端與偵測電極距離不同下之實際電流響應訊號....66
圖3-17：進樣時間的探討........................................................................69
圖3-18：不同進樣時間下之實際電流響應訊號....................................70
圖3-19：探討分離時樣品槽及樣品廢液槽中控制的電壓大小............72
圖3-20：分離時樣品槽及樣品廢液槽中控制電壓改變時的實際電流響應訊號......................................................................................................73
圖3-21：分離電壓的探討........................................................................75
圖3-22：不同分離電壓下之實際電流響應訊號....................................76
圖3-23：網印碳電極偵測acetaminophen與p-aminophenol之校正曲線與實際電流響應訊號..........................................................................78
圖3-24：偵測acetaminophen與p-aminophenol之穩定性......................79
圖3-25：acetaminophen水溶液之半衰期探討........................................80
圖3-26：acetaminophen水溶液之Arrhenius圖.......................................81
表(一) ：偵測系統之最佳化條件與分析特性.......................................83

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