碳奈米粒子是一個非常新興的螢光奈米物質，它具有了許多的特點，像是：容易製備、水溶性佳、化學惰性、低毒性、容易進行表面修飾及抵抗光漂白的能力。因此，在具有這些優勢的特性下，碳量子點能夠應用在許多領域方面，如：光催化、放光裝置、化學感測、金屬離子探針、生化感應器、生物顯影甚至是藥物傳遞方面。
本研究利用聚吡咯修飾之碳奈米粒子（PPy-Cdots）檢測褪黑激素（melatonin）：聚吡咯是一導電高分子，它具有了（1）：生物相容性，可以使環境造成的干擾降低；（2）良好的訊號傳遞，不論是輸入/出訊號，都不易受到本身修飾的影響；（3）：它也可保護結合的材料受到汙染或是干擾，進而影響分析結果。褪黑激素是一種在生物體中普遍存在的激素，它影響了我們人體內的生理時鐘、自然作息，它也具有抗癌及增加免疫的功能。近年來，褪黑激素已經應用在多種疾病治療上，如：失眠及憂鬱症，因此檢驗其在人體的含量是一重要課題。
因此，我們將所合成的碳奈米粒子修飾上聚吡咯高分子，不止使碳奈米粒子的放光能力增加，在與分析物褪黑激素結合時，也能避免其干擾。褪黑激素與所合成的聚吡咯修飾之碳奈米粒子有相似吸收範圍，因此當樣本接受光訊號時，部分的能量會由外在的褪黑激素吸收，使得樣本的放光衰弱，藉由此消光現象來進一步定量出分析物的濃度。

Carbon dots is a very novel nanomaterial with fluorescence property. It has multiple characteristic, such as : easily and facile synthesis, good water solubility, chemistry inert, lower toxic, surface functionalize and resisting photobleaching. Therefore, with all the superioritys, carbon dots can apply in many research regions, like : photocatalysis, photoluminescence, chemosensors, metal ions probes, biosensors, bioimaging even in the drug delivery system.
This thesis is about using polypyrrole functionalize carbon dots(PPy-Cdots) to detect melatonin. Polypyrrole is a conducting polymer, it has three main properties to functionalize it to our synthesis carbon dots. The first one is good biocompatible property that reducing the error from the environment. The second is well signal transferring. No matter what importing or exporting signals, it will not easily influence by functionalize agent itself. The third it can protect carbon dots from polluting and let the analysis result inaccurate. Melatonin is a hormone that is general presence in organisms. It can affect our biological rhyme, natural rest. It also has functions of anticancer and increased immunity. Recently, melatonin is already applied in curing many kinds of diseases, for instance: insomnia and depression. As a result, detecting the quantity of melatonin in human body is an important subject.
As a consequence, we synthesis a polypyrrole functionalize catbon dots, not only make the luminescence increased but also preventing interference from possible resources. Melatonin has similar absorbance region with our sample, so partial energy will absorb by analytes when sample excited by laser cause quenching effect happened. Later, we use this phenomenon to quantify the amount.

目錄
第壹章：總體簡介（General introduction） 1
一、奈米科技的簡介（Introduction of nanotechnology） 1
二、碳奈米粒子簡介（Carbon nanoparticles or Carbon-dots，CDs） 2
三、碳奈米粒子放光特性（Photoluminescent of Carbon nanoparticles） 3
Ⅰ：共軛π軌域能帶躍遷 3
Ⅱ：表面缺陷所導致的放光 4
Ⅲ：可調式的放光 5
Ⅳ：上轉換的放光 6
四、碳奈米粒子的合成方式（Synthesis of carbon nanoparticles） 8
五、參考資料（References） 10
第貳章：材料簡介（Introduction of materials）........................................................13
一、分析物－褪黑激素（melatonin）...................................................................13
二、褪黑激素的偵測方法（Detection methods of melatonin）...........................17
三、修飾材料－聚吡咯（Polypyrrole）................................................................26
四、參考資料（References）................................................................................26
第參章：利用聚吡咯修飾之碳奈米粒子檢測褪黑激素（Carbon dots modified with polypyrrole to detect melatonin.） 28
一、偵測原理（Principle） 28
二、藥品、合成方法及實驗操作（Chemicals、synthesis and procedure of experiments） 28
藥品（Chemicals） 28
合成方法（Synthesis） 29
實驗操作（Procedure of experiments） 30
三、結果與討論（Result and discussion） 32
所合成奈米粒子的物理性質（Physical properties of nanoparticles） 32
利用螢光靈敏度偵測褪黑激素（Melatonin detection using fluorescence） 42
四、結論（Conclusion） 46
五、參考資料（References） 46
第肆章：論文總結（Thesis conclusion）..................................................................51
第伍章：附錄（Appendix）........................................................................................52
圖次
圖1.1：（A）具有強UV吸收但弱或無放光特性的碳奈米粒子，（B）弱UV吸收但具有多色可見光放光特性的碳奈米粒子。 4
圖1.2：（A）PEG1500CDs水溶液在（a）400 nm激發光以濾鏡濾至特定波長激發光及（b）特定波長激發光下的放光圖（B）以PPEI-EI修飾的CDs的吸收及放射光譜圖，激發光以400 nm起（左）以20 nm增量來調節激發光波長。內置圖為放光強度以該CDs的量子產率歸一化後的螢光強度圖。 5
圖1.3：（A）CDs的上轉換放光光譜，激發光以700 nm起（左）以25 nm增量來調節激發光波長。Inset為規一化的放光圖。（B）另一CDs的上轉換放光光譜，激發光從805nm到1035nm。（C）CDs水溶液以800nm的雷射激發的照片。（D）400nm及800nm波長的激發光強度與放光強度的關係圖。 7
圖1.4：碳奈米粒子的合成方式及其後續修飾改質示意圖。 8
圖1.5：利用雷射剝蝕法之碳奈米粒子的一步合成，使用PEG200N溶劑及石墨。 9
圖1.6：利用改質的二氧化矽球體當作基底，苯酚甲醛樹脂作為碳的前驅物的碳奈米粒子合成。 10
圖2.1：褪黑激素在人腦中松果體分泌機制。 13
圖2.2：褪黑激素的功用示意表。 14
圖2.3：人體中合成褪黑激素的路徑表。 15
圖2.4：褪黑激素的歷史時間軸。 16
圖2.5：Total Ion Current圖（TIC圖） 18
圖2.6：（左）褪黑激素濃度與訊號面積的校整曲線，（右）校整曲線的數據表格。 18
圖2.7：褪黑激素與反應後的三種代謝物，（A）褪黑激素MEL，（B）AFMK，（C）AMK，（D）3COHM。 19
圖2.8：褪黑激素與反應後的三種代謝物的（A）吸收色譜圖（B）以210, 231, 279 及380nm激發波長的層析光譜圖。 20
圖2.9：四種分析物：MEL, AFMK, AMK, 5-MT 及3COHM.的HPLC層析圖。 20
圖2.10：褪黑激素及多巴胺在Gr–Fe3O4/GCE電極表面的氧化機制。 22
圖2.11：相似分析物（a）檸檬酸（AA）（b）尿素（UA）的選擇性區分。 22
圖2.12： Gr–Fe3O4/GCE電極對分析物的定量測試（a）定量的多巴胺及變量的褪黑激素，（b）定量的褪黑激素及變量的多巴胺，（c）變量的褪黑激素及多巴胺，（d）c圖的分析物電訊號及濃度檢量線。 23
圖2.13：ELISA實驗流程圖。 24
圖2.14：0.125 μg/mL 至1 μg/mL的褪黑激素抗體吸收曲線。 24
圖2.15：七種褪黑激素的類似物的吸收曲線。 25
圖3.1：利用聚吡咯修飾之碳奈米粒子檢測褪黑激素的原理示意圖。 28
圖3.2：碳奈米粒子（Cdots）合成步驟流程示意圖。 29
圖3.3：聚吡咯（PPy）合成步驟流程及反應原理示意圖。 29
圖3.4：聚吡咯修飾之碳奈米粒子（PPy-Cdots）合成步驟流程示意圖。 30
圖3.5：所合成化合物的紫外光吸收光譜圖。聚吡咯（PPy）、碳奈米粒子（Cdots）、聚吡咯修飾之碳奈米粒子（PPy-Cdots）及褪黑激素（Melatonin）。 33
圖3.6：以不同激發光波長激發（a）碳奈米粒子，（b）聚吡咯修飾的碳奈米粒子之放光光譜圖 35
圖3.7：穿隧式電子顯微鏡及粒子直徑大小分布圖（a）碳奈米粒子，（b）修飾聚吡咯之碳奈米粒子，（c）聚吡咯。 37
圖3.8：所合成化合物的傅立葉轉換紅外線光譜圖，聚吡咯PPy（下），碳奈米粒子Cdots（中），修飾聚吡咯之碳奈米粒子PPy-Cdots（上）。 39
圖3.9：（a）碳奈米粒子及修飾聚吡咯之碳奈米粒子的pH值關係圖，（b）修飾聚吡咯之碳奈米粒子的鹽類穩定性，（c）修飾聚吡咯之碳奈米粒子及結合褪黑激素的複合物（PPy-Cdots-melatonin）之pH值關係圖，（d）PPy-Cdots-melatonin 在pH值6.0～8.0的相對變化圖。 41
圖3.10：修飾聚吡咯之碳奈米粒子在有無褪黑激素存在下的放光特性，將200µL聚吡咯修飾之碳奈米粒子原始溶液，添加至2.8mL的500nM褪黑激素溶液中，以波長360nm的激發光激發樣本，偵測其添加前後放光值的變化。 42
圖3.11：（a）修飾聚吡咯之碳奈米粒子在pH值7.0環境下，對不同濃度的褪黑激素下的螢光靈敏度及（b）以褪黑激素的濃度對上相對放光值的校整曲線關係圖。 43
圖3.12：修飾聚吡咯之碳奈米粒子對於褪黑激素的選擇性測試，分析物濃度為200nM，在pH值7.0的環境下，使用波長360nm的激發光偵測其放光強度。 44
圖3.13：（a）再添加血漿到修飾聚吡咯之碳奈米粒子在pH值為7.0下，對不同濃度的褪黑激素的螢光靈敏度及（b）以褪黑激素的濃度對上相對放光值的校整曲線關係圖。 45
表次
表2.1：HPLC及MEKC方法的多種參數及偵測極限。........................................21
表2.2：使用HPLC及MECK方法偵測5μg/mL的分析物之實際偵測量。.........21
表3.1：本篇研究方法與之前文獻研究比較表。................................ .... .................46

第壹章
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