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研究生：	陳昱翔 Chen, Yu-Hsiang
論文名稱：	以模印高分子修飾電極用於尿液肌酸酐之電位式感測 Preparation of molecularly imprinted polymer modified electrode for the potentiometric detection of creatinine in urine
指導教授：	許梅娟 Syu, Mei-Jywan
學位類別：	碩士 Master
系所名稱：	工學院 - 化學工程學系 Department of Chemical Engineering
論文出版年：	2021
畢業學年度：	109
語文別：	英文
論文頁數：	52
中文關鍵詞：	生醫感測器、肌酸酐、模印高分子、電化學
外文關鍵詞：	biosensor, creatinine, molecularly imprinted polymer (MIP), electrochemistry
相關次數：	點閱：111 下載：0
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近年來隨著社會高齡化的現象，大眾對於衛生保健的意識及重視逐漸抬頭，為了即時監測身體的健康指標，具有快速檢驗、高靈敏度、微量樣本、非侵入、可攜式且使用簡便等優點的生醫感測器成為備受矚目的檢測工具，已能提供即時診斷並預測疾病走向為遠景，希望能達到預防勝於治療的最終目標。
肌酸酐 (Creatinine) 為人體血液與尿液中作為評估腎臟功能的重要指標之一，臨床上常使用傑夫呈色法 (Jaffé reaction)，利用苦味酸 (picric acid) 與肌酸酐之呈色反應進行檢測，其缺點為操作不便且檢測精確度容易受到干擾物影響。為了達到快速且高度精確的檢測，本研究結合電化學與具有高度選擇性的模印高分子 (molecularly imprinted polymer, MIP) 技術，期望能在肌酸酐感測上提供具有可行性與展望性的感測方法。
本研究以肌酸酐作為模版分子，以自行合成的ANVP (2-amino-3-nitro-5-vinylpyridine) 及NVI (1-vinylimidazole) 作為功能性單體進行模印高分子薄膜的合成以修飾金電極表面，透過淬洗過程將肌酸酐從高分子結構中洗脫，留下具有高度辨識性的孔洞作為肌酸酐的吸附位置。透過FTIR之官能基特徵峰存在與否來做為肌酸酐成功洗脫的依據，並以電化學分析各階段修飾電極以進行電化學活性面積與阻抗的探討，在進行感測環境與合成條件的最適化後，利用電位式 (potentiometry) 對肌酸酐作定量檢測。最終在標準尿中於 10300 mg/dL的範圍呈高度線性，且在干擾物與相似物測試的結果中可發現此感測方法具有對肌酸酐的高度專一性，本研究開發之感測器提供了對尿液肌酸酐具有可行性感測的方法。

With the aging of our society in recent years, the publics gradually pay more attention to the health issue. To achieve the concept of point-of-care testing (POCT) and real-time monitoring to the health indicators of our body, development of biosensors with fast response, high sensitivity, high accuracy and non-invasive tends to be more and more important. With the vision of providing immediate diagnosis and further preventing diseases, we hope to achieve the final goal, “Prevention is the better cure.”
Creatinine is one of the most important indices for the evaluation of kidney function. Colorimetry based on Jaffé reaction involves the reaction of creatinine with picric acid under base environment. It is the main measurement for creatinine in clinical diagnosis. However, it is inconvenient to operate and normally suffers from interference.
In this work, the core technique is molecularly imprinting. We combined molecularly imprinted polymer (MIP) technique and electrochemistry for the determination of creatinine in urine. Creatinine as the template molecule, the synthesized ANVP (2-amino-3-nitro-5-vinylpyridine) and NVI (1-vinylimidazole) as the co-monomers, were polymerized onto the gold electrode for the fabrication of the MIP modified electrode. Via the extraction process, creatinine can be removed from the MIP film. Therefore, the cavity specific for recognition and binding of creatinine was created. FTIR, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to analyze the properties of MIP modified electrodes. The as-prepared biosensor shows good linearity in calibration with potential change against creatinine concentration. The calibration of creatinine concentration was in the range of 10 to 300 mg/dL in standard urine and the detection by potentiometric method was applied. The interference test reveals that the proposed MIP biosensor shows sensing specificity towards creatinine molecules. Consequently, it can be concluded that the creatinine biosensor proposed in this work is feasible for the detection of creatinine concentration in urine and also be a promising tool for the applications in clinical diagnosis.

摘要 I
Abstract II
Extended Thesis Abstract III
誌謝 VI
Table of Contents VII
List of Tables X
List of Figures XI
Chapter 1 Introduction 1
1 Preface 1
1.1 Point-of-care testing (POCT) 1
1.2 Biosensor 1
2 Research motivation	2
Chapter 2 Literature review 3
1 Creatinine 3
1.1 Metabolism of creatinine 3
1.2 Creatinine and Kidney disease 3
1.3 Chronic kidney disease and clinical evaluation 4
1.4 Methods/techniques for creatinine determination 4
2 Molecularly imprinted polymer (MIP) 5
2.1 Introduction 5
2.2 Essential components of MIP 6
2.3 Different types of imprinting method 7
2.4 Method of polymerization 8
3 Electrochemical analysis 10
3.1 Introduction 10
3.2 Voltammetry 10
3.3 Chronoamperometry 12
3.4 Electrochemical impedance spectroscopy, EIS 13
3.5 Open circuit potential, OCP 15
Chapter3 Experimental methods, materials and instruments 16
1 Preparation of 2-amino-3-nitro-5-vinylpyridine (ANVP) 16
1.1 Synthesis of ANVP 16
1.2 Purification of ANVP 17
1.3 Thin layer chromatography of ANVP 17
2 Fabrication of creatinine imprinted polymer modified Au electrode 17
2.1 Preparation of thiol modified gold electrode 17
2.2 Preparation of pre-polymerization solution for MIP/NIP 17
2.3 Fabrication of the MIP/NIP modified electrode 17
2.4 Extraction of the MIP/NIP film 18
3 Analysis of electrochemical property 18
3.1 Electrochemical analysis for ANVP 18
3.2 Cyclic voltammetry for the MIP modified electrode with different extraction time 19
4 Optimization of conditions 19
4.1 Optimization of pre-balance time 19
4.2 Optimization of ratio of functional monomer to crosslinker 19
4.3 Optimization of the volume of pre-polymerization solution 19
5 Application tests 19
5.1 Interference test for species in urine 19
5.2 Interference test for analog compounds 19
5.3 Reproducibility test 20
6 Chemicals 21
7 Instruments 22
Chapter4 Results and Discussion 23
1 Functional monomer 23
1.1 Preparation of ANVP 23
1.2 Purification of ANVP 23
1.3 1H-NMR analysis of ANVP 24
1.4 FTIR analysis of ABNP and ANVP 25
1.5 Electrochemical property of ANVP 25
2 Extraction of the molecularly imprinted polymer 26
2.1 Electrochemical analysis of different extraction time 27
2.2 SEM analysis of the MIP film before/after extraction 27
2.3 FTIR analysis of the MIP film before/after extraction 28
3 Modification of the MIP electrode and the electrochemical behavior 29
3.1 Electrochemical property of pre-polymerization solution 29
3.2 Analysis of electrode surface area 29
3.3 Analysis of electrochemical impedance spectroscopy 32
3.4 Signal response by open circuit potential 34
4 Optimization of detection/synthesis condition 34
4.1 Pre-balanced time for detection 34
4.2 Surface modification of gold electrode surface 35
4.3 Ratio of functional monomer to crosslinker 36
4.4 Volume of pre-polymerization solution 37
5 Molecularly imprinted polymer & non-molecularly imprinted polymer 38
5.1 Non-molecularly imprinted polymer, NIP 38
5.2 Imprinting factor 39
5.3 Analytical performance between MIP and NIP 39
6 Application tests of MIP electrode 39
6.1 Interference test for species in urine 40
6.2 Interference test for analog compound 42
6.3 Reproducibility test 43
7 Detection of creatinine in standard urine 43
7.1 Response time 43
7.2 Calibration curve of creatinine 44
7.3 Limit of detection (LOD) and limit of quantification (LOQ) 45
8 Comparison this research to previous literature 46
Chapter 5 Conclusion 48
References 49
                                    

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2026-08-26公開

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