臺灣博碩士論文加值系統

皮質醇普遍稱為逆境激素，其變異或異常被歸因於疾病與綜合症狀，因此皮質醇檢測對於篩檢與監測各種健康狀況是極為重要的。我們的工作使用電化學表面增強拉曼散射（EC-SERS）法開發Au @ PPy核/殼NP，用於檢測唾液中的皮質醇。AuNPs運用R6G於評估SERS的增強因子，其塗佈於平均大小為46nm的PPy殼厚度，其PPy殼厚度取決於Py的聚合時間與單體體積。Au @ PPy（〜46 nm核心/〜1 nm外殼厚度）可獲得的最高增強因子高達1.74 x 106。我們選擇其條件去結合抗體（CMab）與EC-SERS的應用。 EC-SERS探針源自抗體-抗原相互作用，從而增加了皮質醇檢測的選擇性與靈敏度，結果指出使用 EC-SERS法的Au @ PPy-CMab可在有限且寬廣的放範圍下6.69 x 10-12 M偵測皮質醇，並可用於檢測人體唾液中的皮質醇。我們的工作提出SERS探針的快速製造法，且可應用於天然基質中監測疾病與綜合症狀進展的感測方法。

Cortisol is commonly called the stress hormone. Its variations or abnormalities might give rise to diseases and syndromes. Thus, detecting cortisol with high sensitivity and confidence is very important for diagnosis and in monitoring various health conditions. This work is to develop Au@PPy core/shell NPs with the electrochemical surface-enhancement Raman scattering (EC-SERS) methods for the detection of cortisol in saliva. The synthesized AuNPs with an average size of ~46 nm, coated with various PPy (polypyrrole) shell thickness by varying polymerization time and volume of Py (pyrrole) monomer, are used to evaluate the SERS enhancement factor with R6G. The highest enhancement factor was up to 1.74 x 106 attained with Au@PPy (~46 nm core/~1 nm shell thickness). The coated nanoparticles were conjugated with antibody (CMab) and further applied for EC-SERS. The interaction between antibody-antigen increases the selectivity and sensitivity of the EC-SERS probes for detecting the cortisol. Cortisol can be successfully detected in a wide range of concentrations by the application of Au@PPy-CMab and EC-SERS method, with a limit of detection (LOD) down to 6.69 x 10-12 M. Cortisol levels in human saliva were also tested for verification. Our work presents a rapid fabrication of SERS probes and can be applied as sensor devices for monitoring disease and syndromes progression in natural matrices.

摘要 I
ABSTRACT III
ACKNOWLEDGMENTS V
TABLE OF CONTENTS VII
LIST OF FIGURES XI
LIST OF TABLES XVII
LIST OF EQUATIONS XIX
LIST OF ABBREVIATIONS XXI
CHAPTER 1: INTRODUCTION 1
1.1 Background of Raman spectroscopy 1
1.2 Principles of Raman Spectroscopy (RS) 1
1.3 Surface Enhanced Raman Spectroscopy (SERS) 3
1.3.1 SERS enhancement mechanism 5
1.3.1.1 Electromagnetic Enhancement mechanism (EE) 8
1.3.1.2 Chemical Enhancement mechanism (CE) 11
1.3.2 SERS Enhancement Factor (EF) 13
1.4 Electrochemical Surface Enhanced Raman Spectroscopy (EC-SERS) 14
1.4.1 History of EC-SERS 14
1.4.2 Theory of EC-SERS 15
1.4.3 Advantage of EC-SERS 16
1.4.4 Applications of EC-SERS in biosensor 19
1.5 Biocompatibility of Au nanoparticles 20
1.6 Principle of shell-isolated nanoparticle enhanced Raman spectroscopy 20
1.7 Cortisol and bioassay method 21
CHAPTER 2: ADVANTAGES AND CHALLENGES OF STUDY 25
2.1 The purpose of EC-SERS 25
2.2 Cortisol detection 25
2.3 Advantages of AuNPs 26
2.4 Advantages of polypyrrole (PPy) coating 27
2.5 Motivation and objectives of the study 27
2.5.1 Motivation 27
2.5.2 Objectives 28
CHAPTER 3: EXPERIMENTAL SECTION AND CHARACTERIZATION 31
3.1 General experimental section 31
3.1.1 Chemicals and reagents 32
3.1.2 Synthesis of gold nanoparticles (AuNPs) 33
3.2 Synthesis of Au@PPy core/shell nanoparticles 34
3.3 Antibody conjugation with Au@PPy core/shell nanoparticles 34
3.4 Preparation of cortisol standard solution 35
3.5 Saliva sample collection 36
3.6 Preparation of SERS and EC-SERS substrates 36
3.7 Characterization techniques 38
3.7.1 The size distribution of AuNPs 38
3.7.2 Ultraviolet-visible absorption spectroscopy (UV-Vis) 38
3.7.3 Scanning Electron Microscopy (SEM) and Energy-Dispersive X-ray Spectroscopy (EDX) measurement 38
3.7.4 Transmission Electron Microscopy (TEM) measurement 38
3.7.5 X-Ray Diffraction (XRD) 39
3.7.6 Fourier Transform Infrared Spectroscopy (FT-IR) 39
3.7.7 SERS measurements 39
3.7.8 Cyclic voltammetry (CV) 39
3.7.9 EC-SERS measurement 40
3.7.10 X-ray Absorption Spectroscopy (XAS) 40
CHAPTER 4: RESULTS AND DISCUSSION 41
4.1 Morphology, optical absorption properties and effect of the shell thickness on the SERS signal of Au@PPy core/shell nanoparticles 41
4.1.1 AuNPs 41
4.1.2 Au@PPy 42
4.1.2.1 PPy coating with different polymerization time 42
4.1.2.2 PPy coating with different volume of Py monomer 48
4.2 SERS spectra of cortisol on the Au@PPy-CMab nanoparticles 56
4.3 The electro-active behavior of Au@PPy-CMab nanoparticles 63
4.4 Effect of potential on the EC-SERS signal 64
4.5 EC-SERS spectra of cortisol on the Au@PPy-CMab nanoparticles 66
4.6 The reproducibility of Au@PPy-CMab 72
CHAPTER 5: CONCLUSIONS AND FUTURE OUTLOOK 75
5.1 Conclusions 75
5.2 Future outlook 75
REFERENCE: 77
APPENDIX: 85

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