肌酸酐 (Creatinine) 是肌酸代謝最終產物，為臨床診斷腎功能重要指標，因此血液及尿液中的肌酸酐濃度是相當重要的生理檢測項目。本研究以共功能性單體製備肌酸酐模版高分子膜披覆於電極上，並以交流阻抗式感測肌酸酐濃度。本研究先合成主要的功能性單體。以 2-amino-5-bromo-3-nitropyridine (ANBP) 為反應物，用乙烯基取代溴，合成單體 2-amino-3-nitro-5-vinylpyridine (ANVP)，再以 1H-NMR進行鑑定，確認單體正確合成。肌酸酐模版高分子薄膜之製備，乃是以肌酸酐為模版分子，加入兩種以上的功能性單體，以共單體進行合成，是由自行合成的ANVP 與 methacrylic acid (MAA)、1-vinylimidazole (1-VD) 及 4-vinylpyridine (4-VP) 等作不同的單體組合，接著加入交聯劑 ethylene glycol dimethacrylate (EGDMA)，在起始劑 2,2′-azobisisobutyronitrile (AIBN) 作用下進行聚合反應，以製備分子模版高分子 (molecularly imprinted polymer, MIP)。
本論文以交流阻抗進行連續式肌酸酐濃度感測，也探討預聚合液中各成份之比例對感測靈敏度及模印指數之影響。由實驗結果可知，功能性共單體與交聯劑之莫爾比為 1: 25 時，電極會有較佳之模印效果。MIP 電極之靈敏度以阻抗與相位角進行估算，分別為 0.68 ± 0.04 ((|Z|–|Zo|)/|Zo| / (mg/dL)) 與 0.41 ± 0.09 ((θ–θo)/θo / (mg/dL))，其阻抗與相位角分析之模印指數分別為 1.94 及 1.78。在肌酸酐選擇性實驗上，是以肌酸 (creatine) 與N-烴基丁二醯亞氨 (N-hydroxysuccinimide, NHS) 分別作為共存物及相似物。在雙成份混合液下共存物之選擇率 (creatinine/creatine)，為 76.52 (以阻抗計)；在雙成份混合液中，以阻抗變化進行估算，則相似物之選擇率 (creatinine/NHS) 分別為 7.49 (各成份濃度10 mg/dL) 及 9.53 (各成份濃度50 mg/dL)。MIP 電極重複使用性測試中，在六次重複使用後，阻抗與相位角回復率仍相當良好；另外，在重複暨保存性方面，經過二十天保存及十次使用後，其穩定性雖隨時間越長而較差，但仍具有辨識能力。綜觀以上，此模版高分子膜電極對肌酸酐具有良好辨識能力，未來能應用於臨床檢測，可更進一步朝微小化生醫感測器發展。

A novel AC impedance sensor based on molecularly imprinted polymer (MIP) has been developed for the measurement of creatinine concentration. Creatinine is a significant biomarker for the diagnosis of kidney function because it can be easily measured in urine and serum. Therefore, the measurement of creatinine became critical in clinical analysis. In this study, we used three combinations of monomers, including 2-amino-5-bromo-3-nitro-pyridine (ANVP), methacrylic acid (MAA), 1-vinylimidazole (1-VD), 4-vinylpyridine (4-VP), as functional co-monomers. In the presence of creatinine template, EGDMA (ethylene glycol dimethacrylate) as the crosslinker was added. With the initiator AIBN (2, 2′-azobisisobutyronitrile) and UV irradiation, photo-polymerization was triggered. The MIP was extracted to remove, then creatinine imprinted polymer was obtained. The results indicate sensitivities of MIP film in impedance and phase angle are calculated by 0.68 ± 0.04 ((|Z|–|Zo|)/|Zo| / (mg/dL)) and 0.41 ± 0.09 ((θ–θo)/θo / (mg/dL)), then imprinting factors in impedance and phase angle analysis are 1.94 and 1.78. The MIP electrode has excellent selectivity tests for creatinine in the presence of creatine and N-hydroxysuccinimide (NHS). Selectivity in impedance analysis for creatinine in the presence of creatine is 76.52. Additionally, selectivities in impedance analysis for creatinine/NHS are 7.49 (10 mg/dL) and 9.53 (50 mg/dL) respectively. In summary, this MIP electrode has great potential in the development of microbiosensor and for being used on clinical detection in the future.

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