均勻結構之複合式鉑銥奈米金屬觸媒/ 葡萄糖氧化酵素電極及其製備與應用

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研究生：	葉旻鑫 Min-Hsin Yeh
論文名稱：	均勻結構之複合式鉑銥奈米金屬觸媒/ 葡萄糖氧化酵素電極及其製備與應用 Fabrication of Homogeneously-structured PtIr Bimetallic Nano-Catalyst/Glucose Oxidase Composite Electrode and its Applications
指導教授：	黃炳照 Bing-Joe Hwang
口試委員:	周澤川 Tse-Chuan Chou 何國川 Kuo-Chuan Ho 王孟菊 Meng-Jiy Wang
學位類別：	碩士 Master
系所名稱：	工程學院 - 化學工程系 Department of Chemical Engineering
論文出版年：	2009
畢業學年度：	97
語文別：	中文
論文頁數：	257
中文關鍵詞：	鉑銥奈米金屬觸媒、雙氧水氧化反應、均勻複合式觸媒/酵素結構、電泳沉積法、微型感測器
外文關鍵詞：	bimetallic nanocatalysts PtIr/C, hydrogen peroxide oxidation reaction (HOPR), homogeneous catalyst/enzyme composite structure, electrophoresis deposition (EPD), mini sensor
相關次數：	點閱：295 下載：9
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上一筆

本研究主要分為兩部分:開發新穎鉑銥奈米金屬觸媒應用於催化雙氧水氧化反應與應用電泳沉積法製備均勻複合式觸媒/酵素結構生物感測器。
由XRD與TEM探討觸媒結構與粒徑大小及分佈並利用電化學測量觸媒對雙氧水之催化活性；應用同步輻射光源之X光原子吸收光譜(XAS)與密度泛含理論(DFT)解釋銥原子加入可修飾鉑原子之d層電子結構與增加觸媒表面官能基(-OH)，能夠有效提高催化雙氧水氧化反應能力。本研究更進一步提出雙氧水氧化反應於觸媒表面之可能反應路徑，提出雙氧水氧化反應速率決定步驟為雙氧水於觸媒表面進行去質子化反應。
另外，本研究中提出一種新穎的均勻複合式觸媒/酵素結構之概念並藉由電泳沉積法同時沉積觸媒材料-鉑銥奈米金屬觸媒與酵素分子-葡萄糖氧化酵素於微型感測器之工作電極表面；由ESCA縱深分析電極結構證實利用電泳沉積法同時沉積觸媒與酵素能夠形成一層均勻結構之複合式觸媒/酵素薄膜。由長時間穩定性測試與酵素催化動力學探討得知均勻複合式觸媒/酵素結構之生物感測器具有長達25天以上的保存期限且Michaelis constant = 5.68 mM，顯示其均勻複合觸媒/酵素結構能提供穩定、三維空間結構之環境使其酵素穩定性與親合性提高並縮短其經由酵素催化之雙氧水與觸媒進行電化學反應之路徑。
本研究針對其製程參數與流程進行詳細的討論與分析後，所製備之均勻結構複合式微型感測器於施加電位0.4 V (vs. Printed Ag/AgCl)下，偵測葡萄糖之感測線性範圍介於2 mM ~ 20 mM，最低偵測極限為0.1 mM，偵測靈敏度為 2.89 μA/mM．cm2(R2=0.995, R.S.D. =3.26%, N=3)；顯示藉由電泳沉積法能應用於製備具有良好再現性、穩定性與準確性之均勻複合式觸媒/酵素結構微型生物感測器。

This investigation mainly consists of two topics: (a) development of novel, bimetallic nanocatalyst PtIr/C and employing nanocatalyst in hydrogen peroxide oxidation reaction (HOPR). (b) Fabrication of the mini-biosensor with the homogeneous catalyst/enzyme composite structure by electrophoresis deposition (EPD) method.
The crystalline and particle size of nanocatalyst were investigated by XRD and TEM, respectively. The catalytic activity was obtained by the amperometric determination of HOPR. The studies of synchrotron based- X ray absorption spectroscopy (XAS) and density functional theory (DFT) calculation demonstrated that the addition of Ir atom modify the d band electronic configuration of Pt atom and enhance the nanocatalyst functionality, consequently promote the HOPR activity. Furthermore, the HOPR mechanism on the catalyst surface has been proposed and the “deprotonation“step was considered to be rate determining step via this investigation.
Moreover, EPD method has been employed to simultaneously deposit the nanocatalyst and enzyme onto the electrode surface. The depth profile analysis of ESCA provided the evidences that EPD method enables to create the homogeneous nanocatalyst/enzyme composite domain. The long term stability and the low value of Michaelis-Menten constant ( Kmapp =5.68 mM ) revealed that the composite matrix provide a stable and three dimensions structure.
After the parameter optimization, the fabricated mini-biosensor showed a linear detection of glucose ranges from 2 mM to 20 mM with a detection limit of 0.1 mM and the maximal sensitivity of 2.89 μA/mM．cm2 (R2=0.995, R.S.D. =3.26%, N=3). Overall, EPD method has been used for fabricating the homogeneous nanocatalyst/enzyme composite mini-biosensor with favorable reproducibility, stability and accuracy.

目錄
摘要	I
Abstract	II
致謝	III
目錄	V
圖引索	X
表引索	XVIII

第一章 緒論	1
1.	前言	1
2.	葡萄糖生物感測器之發展	2
3.	研究動機與目的	14
第二章 理論基礎與文獻回顧	17
1.	感測器簡介	17
2.	生物感測器簡介	18
2.1.	生物感測器之定義	18
2.2.	生物感測器之特點	18
2.3.	生物感測器之基本構造與原理	19
2.4.	生物感測元件	21
2.5.	信號轉能器	25
3.	酵素感測器	28
3.1.	酵素特性	28
3.2.	酵素單位	28
3.3.	酵素的分類	28
3.4.	酵素專一性反應	30
3.5.	等電點	30
3.6.	葡萄糖氧化酵素	31
4.	電化學生物感測器	33
4.1.	電位式（Potentiometric）生物感測器	34
4.2.	電流式（Amperometric）生物感測器	34
4.3.	電導式 (Conductometric)生物感測器	36
5.	生物分子固定化技術[41]	37
5.1.	生物分子固定化之簡介	37
5.2.	生物分子固定化之擔體	38
5.3.	生物分子固定化之方法	38
5.4.	吸附法 (Adsorption)	42
5.5.	膠囊法 (Encapsulation)	42
5.6.	包埋法 (Entrapment)	42
5.7.	化學鍵結法 (Covalent attachment)	43
5.8.	交連架橋法 (Cross-linking)	43
6.	電泳沉積法(Electrophoretic Deposition)	44
6.1.	電泳沉積法之演進	44
6.2.	電泳沉積法之原理	45
6.3.	電泳沉積法之動力學	48
6.4.	電泳沉積法之文獻回顧	49
6.5.	影響電泳沉積法之參數	50
7.	奈米粒子	50
7.1.	奈米粒子之簡介	50
7.2.	奈米粒子之製備方法	51
7.3.	奈米粒子在生物感測器之文獻回顧	52
第三章 實驗設備與方法	54
1.	實驗藥品與樣品配置	54
1.1.	實驗藥品	54
1.2.	樣品配製	55
2.	實驗設備	56
3.	微型感測器(Mini Sensor)	56
4.	實驗方法	59
4.1.	奈米金屬觸媒之合成方法	59
4.1.1.	擔體前處理	60
4.1.2.	NaBH4還原方法合成Pt/C觸媒	60
4.1.3.	NaBH4還原方法合成Ir/C觸媒	61
4.1.4.	NaBH4還原方法合成Pt1Ir1/C 觸媒	62
4.1.5.   修飾Watanabe法合成Pt1Ir1/C/C-Watanabe觸媒	63
4.1.6.	實驗架構	65
4.2.	電泳沉積法沉積奈米金屬觸媒與葡萄糖氧化酵素於微型感測器之流程		66
4.2.1.	電泳溶液之製備	66
4.2.2.	電泳沉積之流程	66
4.2.3.	實驗架構	67
5.	分析儀器與方法	71
5.1.	觸媒元素鑑定與結構分析	71
5.1.1.	X光繞射分析(XRD)	71
5.1.2.	穿透式電子顯微鏡分析(TEM)	72
5.1.3. 感應偶合電漿放射光譜儀分析(ICP-AES)	   		73
5.2.	微型感測器表面結構分析	73
5.2.1.	化學電子能譜分析儀(ESCA)	73
5.2.2.	掃描式電子顯微鏡分析(SEM)	73
5.3.	電泳溶液之物性分析	74
5.3.1.	表面電位分析儀	74
5.3.2.	導電計	75
5.4.	電化學分析原理	75
5.4.1.	旋轉電極之電化學測試系統	75
5.4.2.	電極製備	76
5.4.3.	旋轉盤電極	76
5.4.4.	微型感測器之電化學測試系統	78
5.4.5.	循環伏安法（Cyclic Voltammetry）	79
5.4.6.	電流應答法 (Amperometric Method)	82
第四章 結果	83
1.	PtIr奈米金屬觸媒之材料結構與電化學特性分析	83
1.1.	金屬觸媒之材料結構分析	84
1.1.1.	ICP-AES分析	84
1.1.2.	XRD 分析	85
1.1.3.	TEM分析	86
1.1.4.	循環伏安分析法	88
1.2.	金屬觸媒之電化學特性分析	90
1.2.1.	循環伏安分析	90
1.2.2.	滴定測試分析法(Titration Method)	94
1.3.	理論模擬計算分析	98
1.3.1.	Pt (111)表面	99
1.3.2.	電荷轉移(charge transfer)	102
1.3.3.	雙氧水於Pt(111)、Ir(111)、PtIr(111)之反應	109
2.	藉由電泳沉積法沉積PtIr奈米金屬觸媒於微型感測器之製程參數與電化學特性分析	114
2.1.	電泳沉積法沉積PtIr奈米金屬觸媒於微型感測器之製程參數			118
2.2.	電泳沉積法沉積PtIr奈米金屬觸媒製備微型感測器之電化學特性分析		120
2.2.1.	電化學活性面積	120
2.2.2.	掃描速率效應	122
2.2.3.	循環伏安分析法	124
2.2.4.	定電位分析法	125
2.2.5.	微型感測器之再現性	127
3.	藉由電泳沉積法同時沉積PtIr奈米金屬觸媒與葡萄糖氧化酵素於微型感測器	129
3.1.	同時沉積PtIr奈米金屬觸媒與葡萄糖氧化酵素於微型感測器之製程參數	129
3.2.	藉由電泳沉積法同時沉積PtIr奈米金屬觸媒與葡萄糖氧化酵素於微型感測器之電極結構分析	131
3.2.1.	微型感測器之表面結構分析	134
3.2.2.	微型感測器之工作電極元素縱深分佈	139
3.3.	同時沉積PtIr奈米金屬觸媒與葡萄糖氧化酵素於微型感測器之電化學特性分析	143
3.3.1.	循環伏安分析法	143
3.3.2.	掃描速率效應	144
3.3.3.	最適化偵測葡萄糖之施加電位	146
3.3.4.	干擾測試	152
3.3.5.	偵測葡萄糖之最低偵測極限	155
3.3.6.	微型感測器之再現性與穩定性	157
3.3.7.	酵素催化反應動力學	160
3.3.8.	微型感測器之長時間保存測試	164
第五章 綜合討論	166
1.	奈米金屬觸媒之材料結構探討	166
2.	奈米金屬觸媒之電化學特性探討	167
3.	奈米金屬觸媒之模擬計算探討	170
4.	藉由電泳沉積法沉積PtIr奈米金屬觸媒於微型感測器之探討	171
5.	藉由電泳沉積法同時沉積PtIr奈米金屬觸媒與葡萄糖氧化酵素於微型感測器之探討	173
第六章 結論	177
第七章 未來方向	179

附錄	180
Appendix A	微型感測器之前處理的參數探討與最適化之研究	180
Appendix A.1.	不同前處理方法之比較	182
Appendix A.2.	利用定電流法前處理微型感測器-時間之影響	187
Appendix A.3.	  利用定電流法前處理微型感測器-電流強度之影響	192
Appendix B   電泳沉積法沉積PtIr奈米金屬觸媒於微型感測器之製程參數	199
Appendix B.1.	電泳溶液之狀態	201
Appendix B.1.1.	 電泳溶液中溶劑之比較	201
Appendix B.1.2.	電泳溶液中Nafion含量之影響	208
Appendix B.1.3.	電泳溶液中觸媒含量之影響	217
Appendix B.2.	電泳程序的條件	219
Appendix B.2.1.	定電壓法之最適化施加電壓與時間	220
Appendix B.2.2.	定電流法之最適化施加電壓與時間	228
Appendix C   同時沉積PtIr奈米金屬觸媒與葡萄糖氧化酵素於微型感測器之製程參數		237
Appendix C.1.	電泳溶液中葡萄糖氧化酵素含量之影響	239
Appendix C.2.	藉由電泳沉積法固定葡萄糖氧化酵素的沉積時間之影響	243

文獻回顧	250

                                

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