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研究生：	江凱焩 Chiang, Kai-Peng
論文名稱：	幾丁聚醣/硫酸化軟骨素奈米粒子之製備及其應用 Preparation and application of chitosan/chondroitin sulfate nanoparticles
指導教授：	洪敏雄 Hon, Ming-Hsiung
學位類別：	碩士 Master
系所名稱：	工學院 - 材料科學及工程學系 Department of Materials Science and Engineering
論文出版年：	2006
畢業學年度：	94
語文別：	中文
論文頁數：	87
中文關鍵詞：	幾丁聚醣、藥物釋放、硫酸化軟骨素
外文關鍵詞：	chitosan, chondroitin sulfate, drug release
相關次數：	點閱：85 下載：4
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本研究以自行設計的步階微滴設備，利用陰電性幾丁聚醣(CTS)與陽電性硫酸化軟骨素(ChS)兩種聚電解質藉著彼此間的靜電庫侖力吸引形成CTS-ChS奈米複合粒子。評估其做為5-FU藥物釋放載體的可行性。傅立葉紅外線光譜分析顯示，CTS的NH3與ChS的COO及OSO3產生鍵結，證實此兩種聚電解質以靜電吸引方式形成錯合物。實驗中分別改變CTS-ChS濃度比、反應pH值以及CTS分子量以觀察其對粒徑及界面電位之影響，最後在CTS-ChS濃度比4:3、反應pH=3以及CTS分子量為80 kDa的條件下粒徑大小為145.1±30 nm的複合微粒。

在藥物釋放的應用上，比較5-FU/CTS與ChS混合及CTS與5-FU/ChS混合兩種不同藥物包覆程序對藥物包覆率的影響，顯示後者所得之包覆率較高為42.5±1.1%；比較起始添加藥量對包覆率的影響，顯示起始添加藥量10 mg所得之包覆率最高為70.4±2.1%。此外，從不同反應pH值及不同CTS分子量的實驗發現到反應時的pH值越高或者CTS的分子量越大，所得之5-FU包覆率越高。而藥物釋放的結果則顯示，不論以600 kDa、200 kDa及80 kDa的CTS所合成的5-FU/CTS-ChS複合微粒，在藥物釋放動力學上均類似兩階段的Higuchi釋放模式，且分子量越小，5-FU的釋放率及釋放速率越大。

In this study, chitosan(CTS) and chondroitin sulfate(ChS), two poly-electrolytes, were used to form nanoparticles through electrostatic force by a step dropwise method. These particles were evaluated their application for drug delivery of 5-fluorouacail(5-FU). According to FTIR analysis, there exists the bonding of NH3 of CTS together with COO and OSO3 of ChS in the complexes. It demonstrates that the complexes are formed by electrostatic force of these two poly-electrolytes. Then, it is observed the influence of the concentration ratio of CTS/ChS, reaction pH value and the viscosity molecular weight(Mv) of CTS on the particle size and zeta potential of CTS-ChS particles. It is found that the CTS-ChS particles with average size of 145 nm could be obtained with the concentration ratio of CTS and ChS of 4:3, reacted in pH 3 condition and the Mv of CTS being 80 kDa.

The in-vitro release profiles of the 5-FU are established in phosphate buffer solution (PBS) pH=7.4 at 37℃. Encapsulation efficiency (EE) of drug for different mixing procedure of 5-FU/CTS with ChS or 5-FU/ChS with CTS is compared, the later shows a higher EE up to 42.5±1.1%. The influence of the amount of initial drug on the EE is shown that the highest EE up to 70.4±2.1% could be obtained for the addition of 10 mg. In addition, as the pH value or Mv of CTS increases, the EE of 5-FU would increase. The result of drug release shows that whether the Mv of CTS is 600 kDa, 200 kDa or 80 kDa, 5-FU/CTS-ChS particles follow two steps Higuchi model for the drug release kinetics. Besides, the release rate and release amount of 5-FU increase for the lower Mv of CTS.

摘要.......................................................................I
Abstract..................................................................II
總目錄...................................................................III
圖目錄...................................................................VII
表目錄.....................................................................X

第一章、緒論...............................................................1
1-1 前言...................................................................1
1-2 研究目的...............................................................3
第二章、文獻回顧...........................................................4
2-1 幾丁質與幾丁聚醣.......................................................4
2-1-1 幾丁質與幾丁聚醣的歷史沿革...........................................4
2-1-2 幾丁質與幾丁聚醣之結構...............................................4
2-1-3 幾丁質與幾丁聚醣之特性...............................................5
2-1-3.1 幾丁質於自然界的分佈...............................................5
2-1-3.2 幾丁質與幾丁聚醣之溶解特性.........................................7
2-1-3.3 幾丁聚醣之去乙醯化程度.............................................7
2-1-4 幾丁質及幾丁聚醣之製備...............................................7
2-1-4.1 幾丁質的製備.......................................................7
2-1-4.2 幾丁聚醣的製備.....................................................8
2-1-5 幾丁質與幾丁聚醣的應用...............................................9
2-2 硫酸化軟骨素..........................................................11
2-2-1硫酸化軟骨素之研究及應用.............................................11
2-3 藥物傳輸技術..........................................................13
2-4 五氟尿嘧啶(5-Fluorouracil)(5-FU)......................................15
2-4-1 5-FU之合成..........................................................15
2-4-2 載體包覆5-FU給藥之發展..............................................16
2-5 奈米藥物載體..........................................................17
2-5-1 奈米藥物載體的製備方式..............................................19
2-6 幾丁聚醣於藥物輸送之應用..............................................20
2-7 幾丁聚醣奈米粒子於藥物輸送之應用......................................21
2-8 聚電解質..............................................................23
2-8-1 聚電解質的介紹......................................................23
2-8-2 幾丁聚醣聚電解質錯合物形成原理......................................25
2-8-3 幾丁聚醣聚電解質反應機制之探討......................................27
第三章、實驗材料與方法....................................................30
3-1實驗藥品...............................................................30
3-2 實驗流程..............................................................31
3-3實驗方法...............................................................33
3-3-1製備不同分子量之幾丁聚醣.............................................33
3-3-2幾丁聚醣分子量之測定.................................................33
3-3-3 幾丁聚醣去乙醯度之測定..............................................34
3-3-4 幾丁聚醣-硫酸化軟骨素複合微粒之製備.................................35
3-3-4-1 溶劑選用..........................................................35
3-3-4-2 實驗方法..........................................................35
3-3-5 5-FU/幾丁聚醣-硫酸化軟骨素複合粒子之製備............................36
3-4 材料分析項目..........................................................37
3-4-1 粒徑及界面電位分析..................................................37
3-4-2 傅立葉轉換紅外線光譜儀(FTIR)........................................38
3-4-3 穿透式電子顯微鏡(TEM)...............................................38
3-5 5-FU/幾丁聚醣-硫酸化軟骨素複合粒子藥物釋放系統分析....................38
3-5-1 5-FU於幾丁聚醣-硫酸化軟骨素複合粒子包覆率計算.......................38
3-5-2 5-FU/幾丁聚醣-硫酸化軟骨素複合粒子之體外溶離實驗....................39
第四章、結果與討論........................................................40
4-1幾丁聚醣分子量及去乙醯度之測定.........................................40
4-2幾丁聚醣-硫酸化軟骨素奈米複合粒子之製備................................44
4-2-1 幾丁聚醣-硫酸化軟骨素奈米複合粒子之組成分子結構分析.................44
4-2-2幾丁聚醣-硫酸化軟骨素之濃度比對於其複合粒子形成之影響................46
4-2-2.1粒徑及界面電位分析.................................................46
4-2-2.2表面形貌分析.......................................................47
4-2-3反應pH值對於幾丁聚醣-硫酸化軟骨素複合粒子形成之影響..................52
4-2-3.1粒徑及界面電位分析.................................................52
4-2-3.2表面形貌分析.......................................................53
4-2-4幾丁聚醣分子量對於其複合粒子形成之影響...............................55
4-2-4.1粒徑及界面電位分析.................................................55
4-2-4.2表面形貌分析.......................................................56
4-3 5-FU/幾丁聚醣-硫酸化軟骨素奈米複合粒子之製備..........................62
4-3-1 5-FU/幾丁聚醣-硫酸化軟骨素奈米複合粒子之分子結構分析................62
4-4 5-FU/幾丁聚醣-硫酸化軟骨素複合粒子之藥物釋放系統分析..................64
4-4-1 5-FU之包覆率分析....................................................64
4-4-1.1 5-FU溶於甲醇之檢量線製作..........................................64
4-4-1.2 5-FU/幾丁聚醣-硫酸化軟骨素複合粒子製備方式對於5-FU包覆率之影響....64
4-4-1.3 起始添加之5-FU藥量對於其包覆率之影響..............................65
4-4-1.4 5-FU/幾丁聚醣-硫酸化軟骨素複合粒子合成時之pH值對於5-FU包覆率之影響
        ..................................................................65
4-4-1.5 幾丁聚醣分子量對於複合粒子之5-FU包覆率影響........................65
4-4-2 5-FU之體外釋放分析..................................................71
4-4-2.1 5-FU溶於磷酸鹽緩衝液(PBS)之檢量線製作.............................71
4-4-2.2幾丁聚醣分子量對於其形成之5-FU/CTS-ChS複合粒子之體外釋放影響.......71
4-4-2.3 藥物釋放動力學之探討..............................................72
第五章、結論..............................................................78
參考文獻..................................................................79
誌謝......................................................................87

                                    

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校內：2007-07-26公開
校外：2007-07-26公開

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