Verapamil主要用於治療高血壓、心絞痛、心律不整等疾病。近年來，新的Verapamil治療用途不斷被開發，如躁症和癌症化療輔助藥物，因此在臨床上準確測定Verapamil濃度是非常必須且必要的。目前臨床分析Verapamil濃度的方法主要是使用高效能液相層析 (HPLC) 搭配UV或螢光偵測器，但其缺點為儀器設備成本昂貴、操作複雜和分析時間較久。
Verapamil化學結構及特性都相當穩定，在一般的電化學檢測結果並不具有電化學反應活性。本研究開發出創新的臭氧前處理法，透過打斷Verapamil分子進而造成 Verapamil結構的改變。經臭氧處理後的Verapamil經電化學量測，具有良好的電化學活性。使用光譜和質譜分析方法都驗證了Verapamil結構的改變，質譜分析更進一步推導出其可能的反應機轉。
本研究開發之電化學微晶片是以金奈米組合電極 (GNEE) 作為工作電極，因金奈米電極具有良好的電化學反應催化性、高靈敏度和高選擇性。整體的分析系統架構是利用金奈米組合電極電化學微晶片搭配循環伏安儀做作電化學分析檢測。此系統優於傳統檢測方法之處為所開發之晶片具有體積小、所需樣品量極小、偵測速度快、容易操作和成本低廉等。
此金奈米組合電極電化學微晶片對於Verapamil水溶液濃度的偵測極限可達10 ng/mL，在100 μg/mL到10 ng/mL範圍內更可達到良好的線性。在血清樣本濃度分析方面，即使是1 ng/mL濃度的Verapamil仍具有良好的電化學活性表現，並在1 ng/mL到100 μg/mL濃度範圍內呈現良好的線性。依據本研究結果可知，金奈米電極電化學微晶片對臨床血清藥物濃度之偵測，確實深具實際應用的可行性。

Verapamil is a commonly used medicine for the treatment of supraventricular arrhythmias, angina and hypertension. Recently, some newly developed applications of Verapamil, such as treating hypomania and chemotherapy for cancers, have been reported. Thus, monitoring the concentration of Verapamil accurately is very important. The major clinical analytical methods of Verapamil concentration determination are high performance liquid chromatography (HPLC) with UV or with fluorescence detector. However, these analytical methods have some disadvantages, like expensive instruments, complex operation, and time-consuming etc.
The chemical structure and properties of Verapamil are very stable. The preliminary result of electrochemical analysis doesn’t show any electrochemical activity. In this study, we developed an innovative ozone pre-treatment method to oxidize Verapamil to the smaller molecules and change its structure. Verapamil have excellent electrochemical activity after ozone pre-treatment. The spectroscopy and mass spectrometry show the changes of Verapamil structure. The products of Verapamil treated with ozone are also predicted by mass spectrometry.
The gold nanoelectrode ensemble electrodes (GNEE) are used as working electrode for its good catalytic activity of electrochemical reaction, high sensitivity and high selectivity. The overall experimental framework of this study is microchip with GNEE working electrode accompanied by cyclic voltammetry, an electrochemical analytical instrument. Compared with traditional analytical methods, the system has some advantages such as small size, micro sample volume, easy operation, rapid detection and low cost.
The limit concentration of Verapamil solution for stable detection in the system is 10 ng/mL. A linear dynamic range with a high correlation factor from 10 ng/mL to 100 μg/mL was obtained. For the analysis of serum sample, Verapamil present excellent electrochemical activity at 1 ng/mL. A linear dynamic range with a high correlation factor from 1 ng/mL to 100 μg/mLwas obtained. According to the results, our system for clinical Verapmil concentration analysis has the feasibility of the practical application.

誌謝 ii
中文摘要 iii
英文摘要 iv
目錄 vi
圖目錄 ix
表目錄 x
符號表 xi
縮寫表 xii
壹、緒論 1
1-1 維拉帕米 (Verapamil) 2
1-1.1 Verapamil 臨床治療應用 3
1-1.2 Verapamil 傳統分析方法 6
1-1.2.1 高效能液相層析分析方法 6
1-1.2.2 光譜分析方法 7
1-2 生物檢測晶片 8
1-2.1 奈米組合電極 9
1-2.1.1 奈米組合電極發展背景 9
1-2.1.2 金奈米組合電極 10
1-2.1.3 金奈米組合電極於生物感測應用 11
1-3 電化學分析方法 12
1-3.1 電化學反應概論 12
1-3.2 循環伏安法 13
1-4 臭氧 16
貳、研究動機與目標 18
參、研究材料與方法 21
3-1 金奈米電極製造 21
3-2 金奈米組合電極電化學微晶片設計 24
3-3 金奈米組合電極電化學微晶片製程和組合 25
3-4 電化學量測架構 27
3-5 Verapamil 樣本溶液配製 28
3-6 Verapamil 血清樣本配製 29
3-7 臭氧前處理 30
3-8 光譜分析 31
3-9 質譜分析 31
3-10 循環伏安法分析 31
3-11 Verapamil血清樣本萃取方法 32
3-12 金奈米組合電極電化學微晶片偵測極限分析 33
肆、結果 34
4-1 金奈米組合電極 34
4-2 金奈米組合電極電化學微晶片 35
4-3 金平面電極與奈米金電極量測比較 36
4-4 金奈米組合電極電化學微晶片分析穩定度 38
4-5 以金奈米組合電極電化學微晶片分析 Verapamil 40
4-6 Verapamil經臭氧處理前後之分析比較 41
4-7 最佳臭氧處理時間分析 45
4-8 VPL/O3 水溶液偵測極限分析 48
4-9 血清中 Verapamil 分析 51
伍、討論 55
陸、未來展望 60
柒、參考文獻 61

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