測量汗液成分濃度對疾病早期診斷與日常保健具有重要意義，例如，肌酸酐是慢性腎臟病的生物標誌物，且汗液可以採用無創的方式採集，能夠成為定點照護檢測(Point-of-care testing，POCT)的良好樣本。然而未處理的汗液樣本中含多種生物分子，例如:葡萄糖、尿酸、乳酸等成分，這些成分可能會干擾檢測結果，所以感測器須具備足夠的選擇性才能應用在POCT上，但是目前肌酸酐感測器的發展面臨選擇性不足與靈敏度不夠高的困境，要將肌酸酐感測器應用在POCT上還是具有相當的挑戰性。
本研究的研究重點在開發同時兼具高靈敏度與選擇性，且成本和儀器需求低的肌酸酐感測器。為提高靈敏度，本研究利用氧化還原石墨稀提高多孔電極表面活性位點數目，在電極表面修飾氧化銅顆粒提高電子轉移速率，並且作為肌酸酐催化的觸媒，修飾過後的電極靈敏度為0.30 ± 0.015µA/µM 和2.63 ± 0.097µA/µM，偵測線性範圍為1-100µM和150-600µM，但是僅修飾RGO與CuO的電極容易受到干擾物的存在影響檢測結果，所以需要在電極表面電接枝自行合成的離子液體與Nafion薄膜，接枝離子液體後感測器能夠排除尿酸外其他干擾物的影響，電極靈敏度為0.16 ± 0.002µA/µM和0.73 ± 0.008µA/µM，線性範圍1.5-100µM 和150-600µM，為了避免尿酸影響檢測結果，需要在電極上再修飾一層Nafion薄膜，利用Nafion上帶負電荷的磺酸基團避免帶正電的尿酸分子接近觸媒，修飾後靈敏度為0.09 ± 0.002µA/µM和1.81 ± 0.003µA/µM，線性範圍為1.5-100µM和150-800µM，所有肌酸酐檢測的校正曲線都具有高線性關係(R2 ≥ 0.99)，100-150µM之間電流與濃度的關係偏離線性，濃度取對數值後才會與電流成正比。此外，本研究為降低成本及提高靈敏度，使用多孔網印電極而不是玻碳電極或一般網印電極，並比較多孔網印電極與一般網印電極在電沉積RGO前後的電化學表現，多孔電極在修飾RGO前CV圖氧化峰電流為291.1µA，一般網印電極僅為37.4µA，在電沉積後多孔電極的Ipa也高於一般網印電極，最後本研究對貯存穩定度和再現性進行測試，在室溫下貯存的感測器在經過40天後電流為原本的96.34%，不同電極製作的感測器測試同一濃度樣品時相對誤差僅1.3(RSD=1.3，n=5)，表現出高貯存穩定性與再現性。

The measurement of sweat component concentrations has significant importance for early disease diagnosis and daily healthcare. For example, creatinine serves as a biomarker for chronic kidney disease, and sweat can be collected in a non-invasive technology, making it a valuable specimen for point-of-care testing (POCT). However, untreated sweat samples contain various biomolecules such as glucose, uric acid, lactic acid, and other components. These constituents can potentially interfere with test results. Therefore, sensors must have sufficient selectivity to be applicable in POCT. However, the current development of creatinine sensors faces challenges related to insufficient selectivity and inadequate sensitivity, making the application of creatinine sensors in POCT quite challenging.
The study focuses on developing a creatinine sensor with high sensitivity and selectivity, while maintaining low cost and minimal instrument requirements. To enhance sensitivity, the study utilizes reduced graphene oxide to increase the number of active sites on the porous electrode surface. The electrode surface is modified with oxidized copper particles to improve electron transfer, serving as a catalyst for creatinine catalysis. The modified electrode demonstrates a sensitivity of 0.30 ± 0.015µA/µM and 2.63 ± 0.097µA/µM, with linear ranges of 1-100µM and 150-600µM. However, the detection results of electrodes modified only with RGO and CuO are easily affected by the presence of interfering substances. Therefore, it is necessary to electrochemically graft ionic liquids and Nafion on the electrode. After grafting ionic liquids, the sensor can eliminate the interferences besides uric acid. The electrode sensitivity is 0.16 ± 0.002µA/µM and 0.73 ± 0.008µA/µM, with linear ranges of 1.5-100µM and 150-600µM. To avoid the interference of uric acid on the detection results, an additional layer of Nafion needs to be modified on the electrode. The negatively charged sulfonyl hydroxide groups on Nafion prevent negatively charged uric acid molecules from approaching the catalyst. After modification, the sensitivity is 0.09 ± 0.002µA/µM and 1.81 ± 0.003µA/µM, with linear ranges of 1.5-100µM and 150-800µM. All creatinine detection calibration curves have high linear relationships (R² ≥ 0.99), the relationship between current and concentration deviates from linearity in the range of 100-150 µM. Only when the concentration is logarithmically transformed, does it show a proportional trend with the current.. In addition, this study want to reduce expense and enhance sensitivity by using a porous screen-printed electrode instead of a glassy carbon electrode or a general screen-printed electrode. The electrochemical performance of the porous screen-printed electrode and the general screen-printed electrode before and after the electro-deposition of reduced graphene oxide (RGO) is compared. Prior to RGO modification, the oxidation peak current in the cyclic voltammetry (CV) of the porous electrode is 291.1µA, whereas the general screen-printed electrode is only 37.4µA. After electro-deposition, the current of the porous electrode is also higher than that of the general screen-printed electrode. Finally, this study tests storage stability and reproducibility. The sensor stored at room temperature maintains 96.34% of its original current after 40 days. Sensors produced with different electrodes, when tested with the same concentration of samples, exhibit a relative error of only 1.3 (RSD=1.3, n=5), demonstrating high storage stability and reproducibility.

摘要-----ii
Abstract-----iv
致謝-----vii
目錄-----viii
圖目錄-----xii
表目錄-----xvii
第一章 緒論-----19
1.1 前言-----19
1.2 研究動機與方法-----20
1.3 實驗規劃-----22
第2章 文獻回顧-----23
2.1肌酸酐的代謝及臨床相關性-----23
2.1.1 肌酸酐的產生-----23
2.1.2 肌酸酐的代謝-----24
2.1.3 肌酸酐濃度和腎絲球過濾率關係-----26
2.1.4 腎絲球過濾率計算公式的演進-----27
2.1.5 及早預防慢性腎臟病的重要性-----31
2.2適用於肌酸酐檢測的生物體液-----33
2.2.1 血液與血清-----34
2.2.2 尿液-----34
2.2.3 唾液-----36
2.2.4 汗液-----36
2.2.5 眼淚-----37
2.3檢測肌酸酐的方式-----38
2.3.1 比色法-----38
2.3.2 電化學法-----40
第3章 實驗方法與材料-----52
3.1網印電極製備-----52
3.1.1 製作混有40wt%碳酸鈣的碳漿-----52
3.1.2 網版印刷碳電極-----52
3.1.3 多孔碳電極製備-----52
3.2電極表面修飾-----54
3.2.1 電極清洗-----54
3.2.2 氧化石墨稀修飾-----54
3.2.3 電化學還原氧化石墨稀-----54
3.2.4 氧化銅修飾-----54
3.2.5 離子液體合成-----55
3.2.6 電接枝離子液體-----56
3.2.7 電接枝陽離子交換膜-----56
3.3感測器分析方式-----57
3.3.1 肌酸酐檢測-----57
3.3.2 干擾物測試-----58
3.3.3 溫度敏感度測試-----58
3.3.4 穩定度測試-----59
3.3.5 再現性測試-----59
第4章 結果與討論-----63
4.1電極設計-----63
4.1.1 電極製作方式-----63
4.1.2 電極修飾-----65
4.2多孔電極的電化學表現與表面特徵-----69
4.2.1 電極表面碳酸鈣完全去除所需時間-----69
4.2.2 多孔電極與一般電極的比較-----71
4.3電沉積氧化石墨烯後之電化學表現-----73
4.3.1 氧化石墨烯沉積液的導電度與pH值-----73
4.3.2 修飾氧化石墨稀後電極表面的顯微結構-----74
4.3.3 電沉積氧化石墨烯對電極活性位點與阻抗的影響-----75
4.4電化學還原GO後電極之電化學表現-----78
4.4.1 RGO的表面特徵-----78
4.4.2 還原氧化石墨稀的阻抗與活性位點變化-----78
4.5氧化銅顆粒的電化學修飾-----81
4.5.1 CuO的表面特徵及修飾後電極的阻抗變化-----81
4.6肌酸酐在修飾電極上的電化學行為-----83
4.6.1 肌酸酐濃度與CV氧化電流的關係-----83
4.6.2 感測器修飾離子液體與Nafion後的校正曲線-----87
4.6.3 實驗數據點以Michaelis-Menten方程式進行曲線擬合-----89
4.7干擾測試-----94
4.7.1 離子液體的合成與分子結構鑑定-----94
4.7.2 電接枝離子液體與Nafion的參數優化-----96
4.7.3 人工汗液干擾測試-----103
4.8溫度敏感度、貯存穩定度與再現性測試-----106
4.8.1溫度敏感度測試-----106
4.8.2貯存穩定度測試-----107
4.8.3再現性測試-----111
4.9 結論與未來展望-----113
4.9.1 結論-----113
4.9.2 未來展望-----113
參考文獻-----115

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