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研究生： 錢培宜
Pei-Yi Chien
論文名稱： 碳六十脂肪酵素/三酸甘油酯壓電晶體感測器研製與應用
Preparation and Application of Piezoelectric Crystal Sensor for Triglyceride Based on C60-Lipase Enzyme
指導教授： 施正雄
Shih, Jeng-Shong
學位類別： 碩士
Master
系所名稱： 化學系
Department of Chemistry
論文出版年： 2007
畢業學年度： 95
語文別： 中文
論文頁數： 121
中文關鍵詞： 三酸甘油酯 、脂肪酵素 、碳六十 、壓電晶體
論文種類： 學術論文
相關次數： 點閱：97  下載：10 
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    • 碳六十脂肪酵素/三酸甘油酯壓電晶體感測器研製與應用

    摘要

    本研究合成碳六十-脂肪分解酵素(C60-lipase)，並應用在三酸甘油酯壓電晶體生物感測器。首先，利用塗佈碳六十的石英壓電晶體感測器作為研究碳六十與脂肪分解酵素間作用力，結果顯示酵素與碳六十存在不可逆的化學性吸附現象。且研究顯示碳六十與脂肪分解酵素間作用的最適溫度為32 ℃，最適pH值為7。
    IR光譜圖利用來鑑定合成的不溶性碳六十-脂肪分解酵素(C60-lipase)。然後，使用多孔性的二氧化矽材質濾片固定化碳六十-脂肪分解酵素，這碳六十-脂肪分解酵素濾片顯示可用來催化水解三酸甘油酯溶液，造成pH值的下降。
    本研究組裝一塗佈碳六十/大環胺醚的石英壓電晶體感測器用來吸附三酸甘油酯溶液經碳六十-脂肪分解酵素催化後的產物-脂肪酸，吸附所引起的負載重量增加會造成石英晶體振盪頻率的下降，並由電腦螢幕紀錄整個催化反應的過程。本研究對三乙酸甘油酯、三丁酸甘油酯及混合三酸甘油酯溶液進行偵測，分別探討了濃度、pH值效應及人體常見生物干擾物存在下的催化反應。結果顯示，此系統對三酸甘油酯的偵測具有良好的濃度線性關係及偵測極限。三乙酸甘油酯、三丁酸甘油酯及混合三酸甘油酯溶液其偵測極限依次為13.08 mg/dL、38.32 mg/dL、6.21 mg/dL皆低於人體內三酸甘油酯正常平均值50~150 mg/dL。同時，此利用C60-lipase而塗佈C60-cryptand[2.2]之生化感測器對於三乙酸甘油酯的偵測具有良好的再現性，RSD值為1.7 % (n= 5)。

    Preparation and Application of Piezoelectric Crystal Sensor for Triglyceride Based on C60-Lipase Enzyme

    Abstract

    Immobilized fullerence C60-lipase was synthesized and applied in a piezoelectric crystal biosensor for triglycerides. Firstly, The C60 coated piezoelectric quartz crystal sensor was employed to study the interaction between C60 and lipase. It seemed to indicate that the lipase could be adsorbed on C60 by chemisorption. The optimum temperature of 32 ℃ and the optimum pH of 7 for the chemisorption between the C60 and lipase were found.
    The water insoluble C60-lipase product was synthesized and identified by IR spectrum. The C60-lipase coated SiO2 filter was also prepared. The C60-lipase filter exhibited the catalytic ability for the hydrolysis of the triglycerides resulting in the decrease in pH of solutions.
    The C60/cryptand[2.2] coated piezoelectric crystal biosensor was prepared for fatty acids , the product of the hydrolysis of the triglycerides catalyzed by C60-lipase. The adsorption of fatty acids on quartz crystal resulted in decrease in the oscillating frequency of quartz crystal. And the overall reaction process can be monitored and recorded by computer. The triglycerides e.g. triacetin, tributyrin and the mixed triglycerides were detected by the piezoelectric biosensor in this study. The effects of concentration, pH and the interference of the common biological species in human blood were also investigated. The piezoelectric sensor has exhibited good concentration linear relation and good detection limit for triglycerides. The detecting limits of the triacetin, tribuytrin and the mixed triglycerides were 13.08, 38.32 and 6.21 mg/dL, respectively which are all below the normal average value of the triglycerides in human body. The C60-cryptand[2.2] coated piezoelectric biosensor with C60-lipase also showed good reproducibility for reusage with RSD = 1.7 % (n = 5) for triglycerides.

    目錄 中文摘要 I 英文摘要 II 目錄 IV 圖目錄 VIII 表目錄 XIII 第一章、緒論 1 1-1 碳六十化學 1 1-1-1 碳六十的發現 1 1-1-2碳六十的基本性質 1 1-1-3碳六十的應用 6 1-2 冠狀醚及大環胺醚 9 1-2-1冠狀醚及大環胺醚簡介 9 1-2-2大環胺醚的應用 11 1-3 酵素化學 12 1-3-1酵素的發現 12 1-3-2酵素的命名 13 1-3-3酵素的特性 14 1-3-4固定化酵素(immobilized enzymes) 19 1-3-5 脂肪分解酵素( lipase ) 27 1-4脂質( lipid ) 31 1-4-1 脂質之簡介 31 1-4-2三酸甘油酯( Triacylglycerols ；Triglycerides ) 33 1-4-3三酸甘油酯的偵測 35 1-5 壓電晶體 36 1-5-1 壓電晶體之壓電性 36 1-5-2 石英振盪器 38 1-5-3 石英振盪器的線路 40 1-5-4振盪頻率的量測 42 1-5-5 石英微量天平(Quartz Crystal Microbalance, QCM) 44 1-5-6 液相石英壓電晶體的特性 48 1-5-7石英壓電晶體之應用 50 1-6研究動機 55 第二章、實驗部分 56 2-1實驗原理與目的 56 2-2藥品與器材 56 2-3石英晶片 56 2-3-1石英晶片之處理 56 2-3-2石英銀電極之表面塗佈液 57 2-3-3表面塗佈法 58 2-4石英振盪系統 59 2-5實驗裝置及步驟 60 2-5-1碳六十與酵素間作用力的研究 60 2-5-2碳六十－酵素之合成 62 2-5-3以pH測定儀檢驗碳六十－酵素的活性 63 2-5-4三酸甘油酯壓電生物感測器之研究 64 2-5-4.1 C60-cryptand[2.2]之合成 64 2-5-4.2碳六十－脂肪分解酵素濾片之製作 65 2-5-4.3三酸甘油酯壓電晶體感測系統 66 第三章、實驗結果與討論 67 3-1碳六十與酵素間的作用力 67 3-1-1塗佈C60/PVC石英晶體對酵素的感應 67 3-1-2碳六十與酵素間的吸附作用力 67 3-1-3C60/PVC的塗佈量效應 67 3-1-4酵素溶液的濃度效應 67 3-1-5酵素溶液的溫度效應 68 3-1-6酵素溶液的pH值效應 68 3-2碳六十固定化酵素 75 3-2-1碳六十/脂肪分解酵素的鑑定 75 3-2-2碳六十/脂肪分解酵素的活性 75 3-3碳六十/大環胺醚的鑑定 78 3-4研製三酸甘油酯壓電生化感測器 80 3-4-1三乙酸甘油酯之偵測 80 3-4-1.1 C60-Cryptand[2.2]/PVC塗佈石英晶體之感應情形 80 3-4-1.2C60-Cryptand[2.2]/PVC的塗佈量效應 80 3-4-1.3三乙酸甘油酯之濃度效應 81 3-4-1.4濾片數目(C60-Lipase量)的影響 85 3-4-1.5三乙酸甘油酯之pH值效應 85 3-4-1.6 再現性 88 3-4-1.7 其他生物分子之干擾研究 88 3-4-1.8固定化酵素濾片多次使用之活性探討 88 3-4-2三丁酸甘油酯之偵測 96 3-4-2.1塗佈C60-Cryptand[2.2]/PVC石英晶體之感測情形 96 3-4-2.2 C60-Cryptand[2.2]/PVC的塗佈量效應 96 3-4-2.3三丁酸甘油酯之濃度效應 96 3-4-2.4三丁酸甘油酯之pH值效應 100 3-4-2.5體內干擾物存在下的影響 100 3-4-2.6 不同三酸甘油酯之感應訊號 100 3-4-3三酸甘油酯混合物之偵測 107 3-4-3.1塗佈C60-Cryptand[2.2]/PVC石英晶體之感測情形 107 3-4-3.2 三酸甘油酯濃度效應 107 3-4-3.3 pH值效應 107 3-4-3.4體內干擾物存在下的影響 111 3-4-3.5濾片儲存活性 111 第四章、結論 117 參考文獻 118

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