臺灣博碩士論文加值系統

本研究以幾丁聚醣為材料，甲酸為溶劑，進行均相的化學反應修飾，利用馬來酸酐與幾丁聚醣的胺基反應，以得到馬來酸取代幾丁聚醣。分別利用重量法及紫外線可見光光譜分析可以求出馬來酸酐轉化率及取代度，在固定反應物比例的條件下(幾丁聚醣/馬來酸酐=1/4)，兩種不同方法求出來的結果相吻合。紅外線光譜及固態核磁共振光譜分析證實於室溫反應下，幾丁聚醣確實與馬來酸酐有化學反應，反應在六小時左右達到平衡值；另外在固定24小時反應時間下，轉化率隨著酸酐量的增加而增加，醣環上的馬來酸取代度亦有增加的趨勢，在馬來酸酐加入重量為幾丁聚醣的5倍時，取代度可到達0.45。再加入光起始劑與不同比例的N-異丙基丙烯醯胺單體(馬來酸取代幾丁聚醣的一倍至五倍重)，利用UV光聚合得到幾丁聚醣聚氮-異丙基丙烯醯胺接枝的薄膜，利用重量分析法分別求出轉化率、接枝效率及接枝比值，結果發現轉化率及接枝效率起初隨著單體加入量的增加而增加，之後就不再改變，平衡轉化率及接枝效率值為94%及56%，而接枝比值則隨單體加入量的增加而增加，當單體加入量為馬來酸取代幾丁聚醣的五倍時接枝比值可達2.47。由於反應後產物共同擁有胺基及羧基，亦擁有聚氮-異丙基丙烯醯胺的溫度敏感性質，隨著接枝量的增加，溫度敏感性值越益明顯，同時LCST溫度會下降，因此可藉由控制反應量的不同，令其擁有不同的pH值及溫度敏感性。針對產物作結構分析、熱分析、膨潤比、型態觀察、細胞培養、LCST測定等測試，藉以評估聚氮-異丙基丙烯醯胺接枝幾丁聚醣材料的性質和生醫適用性。

In this study, chitosan was modified with maleic anhydride in formic acid through the reaction of amino group with anhydride group. In order to obtain products with different degree of substitution, different amount of maleic anhydride was employed to react with chitosan. The results showed that the degree of substitution increased with the added amount of maleic anhydeide.The maximum degree of substitution was 0.46 in this study. In addition, the conversion increased with reaction time and reached a plateau value after 6h. After reaction, different amount of N-isopropylacrylamide monomer and photoinitiator were added to prepare PNIPAAm-graft-chitosan copolymer membranes by UV radiation. The maximum graft efficiency was 52.2% in this study. Because the products have amino group and carboxyl group, they have different swelling ratio values in different pH environments. In addition, the copolymer membranes also have thermal sensitive properties due to the grafted PNIPAAm cahins. The pH and the thermal sensitive behaviors of graft copolymers can be controlled by changing the grafting ratio of PNIPAAm to chitosan. FTIR and NMR were used to analyze the chemical structures of copolymers. DSC and TGA were employed to measure the thermal properties of the prepared copolymers. Swelling ratio was measured to understand the pH and thermal responsive behaviors. Finally, cell biocompatibility was evaluated by cell culture on the membranes.

中文摘要 ………………………………………………………… 1
英文摘要 ………………………………………………………… 2
第一章 緒論 …………………………………………………… 3
第二章 文獻回顧 ……………………………………………… 4
2.1 幾丁質與幾丁聚醣 ……………………………………… 4
2.1.1 幾丁質的製備與型態 ……………………………… 6
2.1.2 幾丁聚醣的製備與去乙醯度的控制 ……………… 8
2.2 幾丁聚醣基本性質的測定 ……………………………… 9
2.2.1 分子量的測定 ……………………………………… 9
2.2.2 去乙醯度的測定 …………………………………… 11
2.3 幾丁聚醣的改質 ………………………………………… 15
2.3.1水溶性幾丁聚醣 ……………………………………… 15
2.3.2烷基化幾丁聚醣 ……………………………………… 20
2.3.3幾丁聚醣的表面改質 ………………………………… 23
2.3.4幾丁聚醣的共聚合反應 ……………………………… 23
2.4智慧型高分子水膠概述 …………………………………… 25
2.4.1溫度敏感型高分子 …………………………………… 25
2.4.2酸鹼敏感型高分子 …………………………………… 27
2.4.3離子強度敏感型高分子 ……………………………… 28
2.4.4光敏感型高分子 ……………………………………… 28
2.5幾丁聚醣與聚氮-異丙基丙烯醯胺共聚物 ………………… 29
第三章 實驗 …………………………………………………… 31
3.1 實驗藥品 …………………………………………………… 31
3.2 實驗儀器 …………………………………………………… 33
3.3 實驗步驟 …………………………………………………… 35
3.3.1 幾丁聚醣去乙醯度的測定 …………………………… 35
3.3.2 幾丁聚醣與馬來酸酐的反應 ………………………… 36
3.3.3 改質後幾丁聚醣與聚氮-異丙基丙烯醯胺接枝反應 … 39
3.4 結構鑑定 …………………………………………………… 41
3.4.1傅氏紅外線吸收光譜(FTIR) …………………………… 42
3.4.2 固態13C-NMR光譜 …………………………………… 43
3.5 馬來酸酐取代度的測定 …………………………………… 43
3.5.1 秤重法 ………………………………………………… 43
3.5.2 紫外光/可見光光譜測定 ……………………………… 44
3.6反應動力研究 ……………………………………………… 44
3.7 聚氮-異丙基丙烯醯胺接枝量的測定 ……………………… 46
3.8熱性質分析 ………………………………………………… 46
3.8.1熱重分析 ……………………………………………… 46
3.8.2微差掃描熱分析儀 …………………………………… 46
3.9場放射掃描式電子顯微鏡 ………………………………… 47
3.10 膨潤比測試 ……………………………………………… 47
3.11 纖維母細胞的培養及活性(MTT)測試 …………………… 47
3.11.1 材料前處理 …………………………………………… 47
3.11.2 細胞培養 ……………………………………………… 48
第四章 結果與討論 ……………………………………………… 49
4.1 幾丁聚醣去乙醯度之測定 ………………………………… 49
4.1.1 去乙醯度的計算 ……………………………………… 51
4.2幾丁聚醣與馬來酸酐的反應 ……………………………… 51
4.2.1 樣品之紅外線吸收光譜分析 ………………………… 51
4.2.2樣品之固態13C-NMR光譜分析 ……………………… 58
4.3轉化率(X，%)及取代度(DS，%)的測定 ………………… 62
4.3.1秤重法 ………………………………………………… 62
4.3.2紫外光/可見光光譜定量 ……………………………… 65
4.3.3反應動力研究 …………………………………………… 69
4.4聚氮-異丙基丙烯醯胺的接枝 ……………………………… 70
4.4.1樣品之紅外線吸收光譜(FTIR)分析 …………………… 71
4.4.2樣品之固態13C -NMR光譜分析 ……………………… 72
4.4.3接枝效率及轉化率 ……………………………………… 76
4.5 性質分析 …………………………………………………… 77
4.5.1熱重分析 ……………………………………………… 77
4.5.2微差掃描熱分析 ……………………………………… 80
4.5.3表面型態分析 ………………………………………… 84
4.5.4 膨潤比分析 …………………………………………… 90
4.5.5 細胞相容性 …………………………………………… 100
第五章 結論 …………………………………………………… 101
第六章 參考文獻 ………………………………………………… 104

1. W. G. Malette, P. L. Omaha, U. S. Patent 4532134 (1985).
2. M. N. Tararel, A. Demard, Biomaterials, V14, 928 (1993).
3. G. J. Tsai, Z. Y. Wu, W. H. Su, J. Food Prot., V63, 747-752 (2000).
4. A. Tokoro, M. Kobayashi, N. Tatewaki, K. Suzuki, T. Mikami, Microbiol Immunol, V33, 357-367 (1989).
5. L. A. Hadwiger, B. Fristensky, R. C. Riggleman, Academic Press, 291-302 (1984).
6. S. Chirachanchai, A. Lertworasirikul, W. Tachaboonyakiat, Carbihydrate Polymers, V46, 19-27 (2001).
7. J. P. M. Sardinha, M. H. Gil, J. V. Mercader, A. Montoya, Journal of Immunological Methods, V260, 173-182 (2002).
8. Seham Nagib, K. Inoue, T. Yamaguchi, T. Tamaru, Hydrometallurgy, V51, 73-85(1999).
9. Tanja Becker, Michael Schlaak, Henry Strasdeit, Reactive ＆Functional Polymers, V44, 289-298(2000).
10. E. Guibal, C. Milot, O. Eterradossi, C. Gauffier, A. Domard, Internation Journal of Biological Macromolecules, V24, 49-59(1999).
11. V. M. Ramos, N. M. Rodríguez, M. S. Rodríguez, A. Heras, E. Agullø, Carbohydrate Polymers, V51, 425-429(2003).
12. P. Perugini, I. Genta, F. Pavanetto, B. Conti, S. Scalia, A. Baruffini, Interional Journal of Pharmaceutics, V196, 51-61(2000).
13. T. Sugama, M. Cook, Progress in Organic Coatings, V38, 79-87(2000).
14. 王綺芬, 1989, 蟹殼幾丁質產品理化性質測定及製備方法之研究, 國立台灣大學食品科所碩士論文。
15. 王鈺驩, 1987, 草蝦頭中幾丁質類產品的製備方法、理化性質與應用, 國立台灣大學農業化學研究所碩士論文。
17. Austin, P. R., Brine, C. J., Castle, J. E. and Zikakis, J. P. 1981.Chitin: New facets of research. Science, 212, 15:749.
18. Minke M, Blackwell J. J Mol Biol, 1978；120:167.
19. Blackwell J, Gardner KH, Kolpak FJ, Minke R, Classey WB. ACS Symp Ser 1980；141:315.
20. Seiichi Mima, Masaru Miya, Reikichi Iwamoto, and Susumu Yoshikawa, Journal of Applied Polymer Science, V28, 1909-1917(1983).
21. Maria Terbojevich, Alessandro Cosani, Bonaventura Focher and Enrico Marsano, Carbohydrate Research, V250, 301-314(1993).
22. Arnold C. M. Wu and Wayne A. Bough, Journal of Chromatography, V128, 87-99(1976).
23. Graham K. Moore and George A. F. Roberts, Int. J. Macromol., V2,(1980).
24. Alasdair Baxter, Michael Dillon, K. D. Anthony Taylor, Int. J. Macromol., V14,(1992).
25. M. L. Duarte, M. C. Ferreira, M. R. Marvão, João Rocha, International Journal of Biological Macromolecules, V31, 1-8(2002).
26. K. M. Vàrum, M. W. Anthonsen, H. Grasdalen, O. Smidsrød, Carbohydrate Research, V211, 17-23 (1991).
27. H. Saito, R. Tabeta, K. Ogawa, Progr. Biotechnol, V3, 267 (1987).
28. Hayes, E. R. and Canada, W., U.S. Patent, No. 4619995, 1986.
29. Muzzarelli, R. A. A., Ilari, P. and Petrarulo, M., International Journal of Biological Macromolecules, 177-180(1994).
30. Nudga, L. A., Plisko, E. A. and Danilov, S. N., Journal of General Chemistry of the USSR, 2729-2732(1974).
31. Y. Shigemasa, H. Usui, M. Morimoto, H. Saomoto, Y. Okamoto, S. Minami, H. Sashiwa, Carbohydrate Polymers,V39, 237-243(1999).
32. Hitoshi Sashiwa, Norioki Kawasaki, Atsuyoshi Nakayama, Einosuke Muraki, Noboru Yamamoto, and Sei-ichi Aiba, Green Biotechnology Research Group, 2002.
33. K. M. Vårum, M. H. Ottøy, O. Smidsrød, Carbohydrate Polymers, V46, 89-98(2001).
34. Mohammad R. Kasaai, Joseph Arul, See L. Chin, Gerard Charlet, Journal of Photochemistry and Photobiology A:Chemistry, V120, 201-205(1999).
35. Anthony L. Andrady, Ayako Torikai, and Takahiro Kobatake, Journal of Applied Polymer Science, V62, 1465-1471(1996).
36. Tadashi Uragami, Seigo Kato, Takashi Miyata, Journal of Membrane Science, V124, 203-211(1997).
37. Mansoor M. Amiji, Colloids and SurfacesbB:Biointerfaces, V10, 263-271(1998).
38. Wen-Yen Chiang, Yuan-An Ku, Polymer Degradation and Stability, V76, 281-290(2002).
39. Loo-Teck Ng, James T Guthrie and Yong J Yuan, Polym. Int., V49, 1017-1020(2000).
40. Mu-Rong Yang, Ko-Shao Chen, Jui-Che Tsai, Ching-Cing Tseng, Shuen-Fun Lin, Materials Science and Engineering C, V20, 167-173(2002).
41. D. K. Singh and Alok R. Ray, Journal of Applied Polymer Science, V53, 1115-1121(1994).
42. Dinesh K. Singh, Alok R. Ray, Journal of Membrane Science, V155, 107-112(1999).
43. M.-C. Yang and W.- C. Lin, Journal of polymer research, V9, 135-140(2002).
44. 徐世昌，1998，甲基丙烯酸甲酯多成分系統之無乳化劑乳化聚合反應-聚合程序，動力機構及性質分析，國立台灣大學化學工程系研究所博士論文。
45. Jun Zhang, Youling Yuan, Jian Shen, Sicong Lin, European Polymer Journal, V39, 847-850(2003).
46. Y.H.Bae, T. Okano, S.W. Kim, Journal of Polymer Science: Part B: Polymer Physical.V28, 923-936(1990).
47. H.Feil, Y.H. Bae, J. Feijen, S.W. Kim, Macromolecules, V25, 5528-5530(1992).
48. Xian-Zheng Zhang, Da-Qing Wu, Chih-Chang Chu, Biomaterials, V25, 3793-3805(2004).
49. T. Shimizu, M. Yamato, A. Kikuchi, T. Okano, Tissue Engineering, V7, 141-151(2001).
50. Z. Ding, R.B. Fong, C.J. Long, P.S. Stayton, A.S. Hoffman, Nature, V411, 59-62(2001).
51. Wen-Fu Lee, Shin-Hsun Yen, Journal of Applied Polymer Science, V78, 1604-1611(2000).
52. Xiao-Lin Wang, Jian Huang, Xiu-Zhen Chen, Xue-Hai Yu, Desalination, V146, 337-343(2002).
53. Ryoichi Kishi, Okihiko Hirasa, Hisao Ichijo, Polymer Gels and Networks, V5, 145-151(1997).
54. Anjali Panda, S.B. Manohar, S. Sabharwal, Y.K. Bhardwaj, A.B. Majali, Radiation Physics and Chemistry, V58, 101-110(2000).
55. Liang Liang, Xiangdong Feng, Loni Peurrung, Vish Viswanathan, Journal of Membrane Science, V162, 235-246(1999).
56. Oh Hyeong Kwon, Akihiko Kikuchi, Masayuki Yamato, Teruo Okano, Biomaterials, V24, 1223-1232(2003).
57. Teruo Okano, Noriko Yamada, Minako Okuhara, Hideaki Sakai, Yasuhisa Sakurai, Biomaterials, V16, 297-303(1995).
58. Byeongmoon Jeong, Sung Wan Kim, You Han Bae, Advanced Drug Delivery Review, V54, 37-51(2002).
59. Kazuyuki Nakajima, Shizuyo Honda, Yasuko Nakamura, Fernando López-Redondo, Shinichi Kohsaka, Masayuki Yamato, Akihiko Kikuchi, Teruo Okano, Biomaterials, V22, 1213-1223(2001).
60. M.K. Yoo, Y.K. Sung, Y.M. Lee, C.S. Cho, Polymer, V41, 5713-5719(2000).
61. Wen-Fu Lee, Chin-Hsuan Shieh, Journal of Applied Polymer Science, V71, 221-231(1999).
62. M. Huglin, Y. Liu, L. Velada., Polymer, V38, 5758-5791(1997).
63. A.R. Khare, N.A. Peppas., Biomaterials, V16, 559-567(1997).
64. 胡孝光、陳美瓊, 化工技術, V7, 174-185(1999)。
65. Chia-Fen Lee, Chia-Jen Wen, Wen-Ten Chiu, Journal of Polymer Science: Part A: Polymer Chemistry, V41, 2053-2063(2003).
66. So Yeon Kim, Sung Min Cho, Young Moo Lee, Seon Jeong Kim, Journal of Applied Polymer Science, V78, 1381-1391(2000).
67. Liliana Verestiuc, Claudia Ivanov, Eugen Barbu, John Tsibouklis, International Journal of Pharmaceutics ,V269, 185-194(2004).
68. L. A. Berkovivh, M. P. Tsyurpa, and V. A. Davankov, Journal of Polymer Science, V21, 1281-1287(1983).