臺灣博碩士論文加值系統

分子模版高分子 (Molecularly imprinted polymers；MIP) 乃是藉以欲分析物或稱模版分子 (template) 先與功能性單體 (functional monomer) 形成以鍵結作用力吸引的複合物型態；聚合時，單體圍繞模版分子，同時加入交聯劑 (crosslinker) 將模版分子固定化。經溶劑萃取方式將高分子內之模版分子洗出，以形成多孔性可辨識區域，即可製備出對欲分析物具有優異親和性及特異選擇性之高分子。
本實驗以判定黃疸之指標性分子，膽紅素作為模版分子，以甲基丙烯酸 (Methacrylic acid；MAA) 及乙二醇二甲基丙烯酸酯 (ethyleneglycol dimethacrylate；EGDMA) 分別作為功能性單體及交聯劑，並加入偶氮異二丁腈 (2,2’-azo-bisisobutyronitrile；AIBN) 起始劑進行熱聚合。製備完成之聚合物以EDTA/NaOH萃洗液萃取包埋於內之膽紅素模版分子，以形成具有辨識膽紅素之孔洞。並藉由比表面積測定 (Brunauer-Emmett-Teller analyzer；BET)、掃瞄式電子顯微鏡 (scanning electron microscope；SEM) 分別進行膽紅素模版高分子之物性測定與表面觀察。膽紅素分子極易因光和氧氣而氧化，故首先探討EDTA/NaOH背景溶液及膽紅素溶液之穩定性。
本研究亦針對膽紅素模版高分子於單成份與多成份混合溶液系統中對膽紅素之吸附選擇性進行討論；並對MIP於單成份及雙成份混合溶液中之吸附動力予以探討，由此進一步建立吸附模式，求取其模式參數並予以討論。除此之外，亦探討加入鹽類對MIP吸附膽紅素之影響。MIP於多成份混合溶液系統下對膽紅素及共存物 (黃體酮、睪固酮、膽固醇) 之吸附選擇性亦予以討論，並延伸多成份系統至真實血清溶液中，且建立MIP對膽紅素之吸附檢量線。

Molecularly imprinted polymer (MIP) was prepared by the interaction between the analyte or the template molecule and the functional monomer. During the polymerization, the cross-linker was added to confine the template molecule. Later, the template bound within the MIP was extracted by the utilization of solvent, the cavities as the recognition sites were thus formed. In this way, the MIP being made can have superior affinity and specificity towards the analyte.
In this research, bilirubin, being the index for jaundice, was used as the template molecule. Methacrylic acid (MAA) and ethyleneglycol dimethacrylate (EGDMA) were used as the functional monomer and cross-linker, respectively. 2,2’-azo-bisisobutyronitrile (AIBN) was added to initiate the thermal polymerization. The MIP thus prepared was then washed by EDTA/NaOH solution for the extraction of bilirubin template molecules. Specific surface area analysis (Brunauer-Emmett-Teller analyzer; BET) and scanning electron microscope (SEM) were used to measure MIP’s physical properties as well as to observe its surface morphology. How the EDTA/NaOH solution affected the stability of bilirubin was investigated. The binding specificities of the MIP in the single solution and binary mixture were compared and discussed. The adsorption isotherm as well as the kinetic model was established. The parameters in the model were obtained and compared. The influence of salt for the adsorption performance was also discussed. The binding specificity of the MIP in triple mixture solution was further investigated. In addition, the serum samples were prepared to establish the calibration of bilirubin concentration via the adsorption of the MIPs.

表目錄I
圖目錄II
第一章 緒論1
1-1 生物感測器的種類1
1-2 分子模版高分子發展及理論3
1-3 分子模版高分子之製作4
1-3-1 製作原理4
1-3-2 功能性單體 (Functional monomers)7
1-3-3 交聯劑 (Crosslinker)7
1-3-4 聚合方式8
1-3-5 溶劑9
1-3-6 模版分子之萃取10
1-3-7 分子模版高分子在各領域之應用10
1-4 膽紅素分子模版高分子之製備14
1-4-1 功能性單體-甲基丙烯酸17
1-4-2 交聯劑-乙二醇二甲基丙烯酸酯17
1-4-3 起始劑-偶氮異二丁腈17
1-4-4 模版分子-膽紅素IXα18
1-5 黃疸 (Jaundice) 及膽紅素 (bilirubin) 之認識19
1-5-1 膽紅素之製造與代謝 (Intestinal phase of bilirubin metabolism)19
1-5-2 膽紅素之化學檢驗21
1-6 膽紅素之三維立體構形分析22
1-6-1 膽紅素之光學異構化及光氧化機制23
1-7 研究動機25
第二章 實驗方法與材料27
2-1 膽紅素穩定度實驗27
2-1-1 EDTA/NaOH與ascorbic acid/NaOH之pH穩定性測試27
2-1-2 膽紅素於EDTA/NaOH或ascorbic acid/NaOH背景溶劑下之穩定性比較27
2-1-3 藍光照射對膽紅素之影響27
2-1-4 氧氣對膽紅素之影響27
2-2 膽紅素模版高分子之聚合方法27
2-3 各組成份析方法28
2-3-1 膽紅素濃度定量測定方法及檢量線28
2-3-2 膽綠素及steroids系列之定量測定方法及檢量線29
2-4 膽紅素模版高分子之膽紅素萃出方法34
2-5 離子強度對吸附之影響實驗34
2-5-1 鹽類對模版高分子吸附膽紅素之影響34
2-5-2 增加鹽類濃度對模版高分子吸附膽紅素之影響34
2-6 膽紅素模版高分子之重複使用實驗34
2-7 模版高分子於多成份系統下之選擇性實驗37
2-8 膽紅素和膽綠素之吸附動力實驗37
2-8-1 單一組成溶液下膽紅素和膽綠素之吸附平衡時間37
2-8-2 單一組成溶液下膽紅素和膽綠素之吸附動力實驗38
2-8-3 雙組成溶液下膽紅素和膽綠素之吸附動力實驗38
2-8-4 模版高分子吸附動力之模式建立38
2-9 模版高分子對血清中膽紅素之吸附39
2-9-1 膽紅素於血清中之檢量線39
2-9-2 模版高分子於血清中對膽紅素之吸附平衡時間39
2-9-3 模版高分子於血清中對膽紅素之吸附檢量線39
2-10 以 β-cyclodextrin (β-CD) 為基材製備膽紅素模版高分子對膽紅素之吸附40
2-11 實驗藥品41
2-12 實驗儀器43
第三章 結果與討論44
3-1 膽紅素穩定度實驗44
3-1-1 EDTA/NaOH與ascorbic acid/NaOH之pH穩定性測試44
3-1-2 膽紅素於EDTA/NaOH與ascorbic acid/NaOH背景溶劑中之穩定性46
3-1-3 藍光照射對膽紅素之影響48
3-1-4 氧氣對膽紅素之影響51
3-2 膽紅素模版高分子之物性54
3-2-1 BET54
3-2-2 SEM54
3-3 膽紅素模版高分子之吸附57
3-3-1 膽紅素模版高分子之吸附平衡時間57
3-3-2 膽紅素模版高分子之選擇性57
3-4 比較以不同材質製備膽紅素模版高分子之可行性61
3-4-1 使用 β-cyclodextrin材質製備膽紅素模版高分子61
3-4-2 Poly(β-CD) MIP與PMAA MIP吸附之比較61
3-5 膽紅素分子模版高分子之重複使用65
3-6 離子強度對吸附之影響實驗67
3-6-1 鹽類對MIP吸附膽紅素之影響67
3-6-2 增加鹽類濃度對模版高分子吸附膽紅素之影響68
3-7 MIP於多成份系統下之選擇性實驗70
3-7-1 混合溶液組成簡介71
3-7-2 MIP於混合溶液中對各成份之選擇性73
3-8 模版高分子對血清中膽紅素之吸附77
3-8-1 MIP對血清膽紅素之吸附平衡時間77
3-8-2 MIP於血清中對膽紅素之吸附檢量線77
3-9 膽紅素和膽綠素之吸附動力實驗82
3-9-1 單一組成溶液下膽紅素和膽綠素之平衡吸附時間82
3-9-2 單一組成溶液下膽紅素和膽綠素之吸附動力實驗82
3-9-3 MIP於雙組成溶液下膽紅素和膽綠素之吸附動力實驗88
3-10 吸附動力之建模95
3-10-1 模式之選擇與建立95
3-10-2 MIP於雙成份系統下吸附動力模式之建立99
第四章 重點整理105
第五章 結論107
參考文獻108

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