本研究的主要目的是利用白金電極對白金二價錯離子的獨特催化性質，藉由微電極微小化具低侵入性的優點發展抗癌藥物-順鉑的電化學偵測平台。 
在過去並未有人探討在人體生理條件下順鉑的電化學行為，實驗第一部分將以一系列循環伏安法探討各種條件下包括酸鹼度、工作電極材質、氯離子濃度、迴轉電壓等，對順鉑的電化學特性的影響，由上述實驗結果推論順鉑在白金電極上的反應途徑為軸向氧化(Axial oxidation pathway)，為增進系統重複偵測的再現性，於量測過程需要經過電位處理活化電極表面。故本實驗採用脈衝電壓以活化電極搭配一直流電壓進行電極的前處理程序。在最佳化處理後，連續偵測順鉑八次的再現性由未處理前的44.3%降低為1.2%，顯示電位處理具有高度的穩定性作用。
經電位處理最佳化後，電化學實驗選擇以微差脈衝伏安法 (Differential Pulse Voltammetry)進行抗癌藥物順鉑的偵測，在溶液環境為0.05 M磷酸緩衝溶液，pH7.4內含0.103 M的氯化鈉條件下，脈衝振幅大小50 mV、脈衝時間75 msec、脈衝周期為200 msec、取樣時間為脈衝結束前16.7 msec的操作條件下，其偵測線性範圍從5 μM 至200 μM (R=0.999)，靈敏度為3.383×10-6A/M，本系統的偵測極限為460 nM (S/N=3)，對於20 μM cisplatin重複操作的相對標準偏差為1.78 % (n=20)。

A novel method for the fast detection of cisplatin by platinum microelectrode has been developed. The typical electrochemical behavior of cisplatin in different parameters including pH, concentration of chloride ion, material of working electrode and its surface condition has been investingated by cyclic voltammetry. Subsequently, cisplatin has measured by differential pulse voltammetry (DPV) using a microelectrode with effective radius of 13.6μm. This method involves a potential pretreated process to renew the activity of electrode surface. In contrast to a series of measurements without the pretreatment process, the relative standard deviation (n = 8) could be reducing from 44.3% to 1.2% compared with original process. After careful optimization, the analytical performance of current microelectrode was evaluation in the buffer solution of 0.05M, phosphate buffer pH7.4 containing 0.103M sodium chloride. The linear range of cisplatin is between 5μM to 200μM (R =0.999) with sensitivity of 3.383×10-6A/M. The estimated detection limit of this scheme is about 460 nM (S/N=3) and the relative standard deviation of twenty respective detection measurement of 20 μM cisplatin is 1.78%.

目錄(Contents)
第一章 緒論……………………………………………………………1
1-1 微電極…………………..……………………………………1
1-1-1 微電極簡介……………………………………………………1
1-2 微電極特性… … … … … … … … … … … … … … … … …2
1-2-1 電阻電容效應…….……………………………………………2
1-2-2 高質傳效應………….…………………………………………3
1-2-3 歐姆壓降效應（IR Drop effect）……………………………4
1-3 微電極製作方法……………………………………………5
1-3-1 電蝕刻…………………………………………………………5
1-3-2 薄層微電極封裝法…………………………………………9
1-4 微電極的應用……………………………………………12
1-4-1 掃描式電化學顯微鏡…………………………………………12
1-4-2 動力學量測……………………………………………………14
1-5 抗癌藥物………………………………………………………15
1-5-1 癌症簡介………………………………………………………15
1-5-2 抗癌藥物的分類………………………………………………16
1-5-3 抗癌藥物-順鉑簡介……………………………………………18
1-5-4 順鉑的抗癌機制………………………………………………20
1-5-5 順鉑的副作用…………………………………………………22
1-5-6 順鉑的偵測…………………………………………………24
1-6 順鉑的氧化還原機制…………………………………………26
1-6-1 Axial oxidation pathway …………………………………………26
1-6-2 Equatorial oxidation pathway…………………………………27
1-7 研究目的……………………………………………………28

第二章 實驗
2-1 儀器設備…………………………………………………………29
2-2 藥品………………………………………………………………29
2-3 微電極製作………………………………………………………30
2-4 微電極特性探討…………………………………………………31
2-5 電化學偵測順鉑原理探討………………………………………31
2-5-1 不同酸鹼值下順鉑的電化學行為……………………………31
2-5-2 氯離子效應……………………………………………………31
2-5-3 工作電極材質選擇……………………………………………31
2-5-4 不同迴轉電壓探討……………………………………………31
2-6 電位前處理………………………………………………………32
2-6-1 穩定電壓最佳化………………………………………………32
2-6-2 前處理平衡時間最佳化………………………………………32
2-6-3 負向脈衝前處理最佳化 ………………………………………32
2-6-4 正向脈衝前處理最佳化…………………………………………33
2-6-5 脈衝次數最佳化………………………………………………33
2-6-6 電位前處理比較………………………………………………33
2-7 溶液條件最佳化………………………………………………33
2-7-1 氯離子濃度探討………………………………………………33
2-7-2 緩衝溶液種類探討……………………………………………34
2-7-3 緩衝溶液濃度探討……………………………………………34
2-8 微差脈衝伏安法最佳條件探討……………………………………34
2-9 白金微電極對於順鉑的分析特性………………………………34

第三章 結果與討論
3-1 電化學順鉑偵測原理………………………………………35
3-1-1 不同酸鹼值下順鉑的電化學行為……………………………35
3-1-2 氯離子效應……………………………………………………38
3-1-3 工作電極材質選擇……………………………………………40
3-1-4 不同迴轉電壓探討 …………………………………………41
3-2 微電極特性……………………………………………………43
3-3 電位前處理…………………………………………………45
3-3-1 穩定電壓最佳化………………………………………………47
3-3-2 穩定電壓施加時間最佳化………………………………48
3-3-3 前處理平衡時間最佳化…………………………………49
3-3-4 負向脈衝前處理電壓最佳化………………………………50
3-3-5 正向脈衝前處理電壓最佳化………………………………51
3-3-6 正負脈衝前處理電壓施加時間最佳化……………………52
3-3-7 脈衝次數最佳化……………………………………………54
3-3-8 電位前處理比較……………………………………………55
3-4 溶液條件最佳化…………………………………………56
3-4-1 氯離子濃度探討………………………………………………56
3-4-2 緩衝溶液種類探討…………………………………………57
3-4-3 緩衝溶液濃度探討…………………………………………58
3-5 微差脈衝伏安法最佳條件探討………………………………59
3-5-1 脈衝振幅大小的探討…………………………………………60
3-5-2 脈衝時間的探討………………………………………………62
3-5-3 取樣時間的探討………………………………………………63
3-6 白金微電極對於順鉑的分析特性…………………………64

3-7 結論… … … … … … … … … … … … … … … … … … … …66
參考資料………………………………………………………………67
圖表目錄
圖1-1：不同RG 比薄層電極的電流密度曲線…………………………9
圖1-2：光固化劑PFPEs 製作程序……………………………………11
圖1-3：SECM 雙工作電極擴散層重疊示意圖………………………12
圖1-4：SECM 回饋示意圖以及電流變化……………………………13
圖1-5：順鉑進出細胞分子的機制……………………………………20
圖1-6：順鉑引起細胞死亡的機制……………………………………21
圖1-7：順鉑水解流程…………………………………………………25
圖1-8：白金四配位錯合物Axial 氧化機制示意圖…………………27
圖1-9：白金四配位錯合物Equatorial 氧化機制示意圖……………28
圖2-1：微電極製作裝置以及電極示意圖……………………………30
圖3-1：順鉑的循環伏安圖譜 ………………………………………36
圖3-2：順鉑的循環伏安圖譜 ………………………………………37
圖 3-3：酸鹼度對順鉑的影響…………………………………………37
圖3-4： 氯離子濃度效應……………………………………………39
圖3-5： 氧化電位偏移統計圖………………………………………39
圖3-6： ITO 及Glassy carbon 電極對順鉑的循環伏安圖……….41
圖3-7：迴轉電壓對順鉑的影響……………………………………42
圖3-8：白金微電極的伏安圖譜…………………………………44
圖3-9： SEM 下白金微電極表面輪廓圖……………………………44
圖3-10： 順鉑以微電極操作所得的的微差伏安圖…………………45
圖3-11：前處理電壓波形示意圖…………………………………46

圖3-12：不同穩定電位前處理探討…………………………………47
圖3-13：不同穩定電壓前處理條件的背景電流……………………48
圖3-14：穩定電位前處理施加時間的影響…………………………49
圖3-15：電位前處理平衡時間探討…………………………………50
圖3-16：負脈衝電壓前處理電位探討………………………………51
圖3-17：正脈衝電壓前處理電位探討………………………………52
圖3-18：負脈衝電壓前處理施加時間探討…………………………53
圖3-19：正脈衝電壓前處理施加時間探討…………………………53
圖 3-20：脈衝次數探討………………………………………………54
圖3-21：電位前處理最佳化施加方式………………………………55
圖3-22：電位前處理統計數據比較…………………………………56
圖3-23：氯離子濃度影響…………………………………………57
圖 3-24：緩衝溶液種類影響………………………………………58
圖 3-25：緩衝溶液濃度影響………………………………………59
圖3-26：微差脈衝伏安法電位變化圖……………………………60
圖3-27：脈衝振伏大小對系統分析影響…………………………62
圖3-28：脈衝時間的效應…………………………………………63
圖 3-29：取樣時間的效應…………………………………………64
圖 3-30：校正曲線圖………………………………………………65
圖 3-31：系統再現性………………………………………………65
表1-1 ：各種不同材料的電化學蝕刻的條件…………………………8
表1-2 ：薄層微電極電流公式整理表…………………………………8
表1-3 ：順鉑常用混合療程…………………………………………19

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