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研究生:林偉哲
研究生(外文):Wei-Che Lin
論文名稱:發展新穎噴霧流化床技術製備微奈米球型金雀異黃酮分子模版應用於豆漿中之記憶性固相萃取
論文名稱(外文):The development of novel spray fluidized bed to technology prepare genistein imprinted micro and nano sphere for memory solid-phase extraction from soybean milk
指導教授:周澤川
指導教授(外文):Tse-Chuan Chou
學位類別:碩士
校院名稱:國立成功大學
系所名稱:化學工程學系碩博士班
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:147
中文關鍵詞:豆漿金雀異黃酮分子模版噴霧流體化床分離純化
外文關鍵詞:soybean milkgenisteinmolecular imprinted polymerspray fluidized bedseparation
相關次數:
  • 被引用被引用:1
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本研究目的在於利用分子模版技術,代替傳統的分離純化程序,自豆漿中提煉金雀異黃酮(genistein)。金雀異黃酮是一種植物性化學物質對人體有許多醫療保健上的作用。它具有預防乳癌、預防前列腺癌、降低心血管疾病的罹患、抗氧化和預防骨質疏鬆等作用。使它成為近年來許多研究人員積極研究之物質。

本實驗室結合分子模版、噴霧流體化床、靜電集塵技術發展一新穎噴霧流體化床聚合系統,希望在此研究領域有所突破。此技術簡化傳統分子模版技術繁複的程序。將目標物、功能性單體、交聯劑、起始劑、溶劑混合成均一相後,以噴霧器將聚合液噴出型成氣懸膠液滴。此懸浮於石英管中呈流體化床的氣懸膠聚合液滴以UV光源照射行光聚合反應並且以靜電集塵板收集完成反應之粉體。此實驗裝置的優點是可以得到單一顆粒微奈米尺寸球型金雀異黃酮分子模版而且可以更有效率的完成聚合反應。

本研究以金雀異黃酮為目標分子進行分子模版的製備。以微熱卡計的恆溫滴定方法計算分子間反應熱,再利用批式再吸附測試,找出最適化設計之分子模版組成。比較恆溫滴定以及批式再吸附的結果,乙二醇二甲基,三甲氧基丙烷三甲基丙烯酸酯 (TRIM)以及甲基丙烯酸 (MAA)分別是於被測試的單體中,最適交聯劑與功能性單體。以單體和交聯劑體積比為1:8 完成聚合反應,之後經過洗模板和再吸附測試,金雀異黃酮分子模版顯示對金雀異黃的吸附量506.59μg/g,膜印係數達1.738。由文獻得知,在流體化床內進行流體化的粒子粒徑範圍再幾十到幾百微米之間,本研究利用噴霧流體化床聚合系統之分子模版平均粒徑在雷光散射法粒徑分析儀分析下可達到90nm。在多成份系統下進行吸附測試,金雀異黃酮分子模版之選擇性可達到50.5。實際運用模版於豆漿萃取液之吸附顯示出對金雀異黃酮之吸附量為80.92μg/g,而對大豆黃酮之吸附量為39.77μg/g。代表分子模版於真實樣品溶液內,對金雀異黃酮分子具有吸附能力以及選擇性。
The purpose of this research was to replace the traditional separation process by molecular imprinting technique to separate and purify genistein from soybean milk. Genistein (4',5,7-trihydroxyisoflavone) is a phytochemical group associated with a range of potential health beneficial effects. These include chemoprevention of breast and prostate cancers, cardiovascular disease, antioxidant, and anti-osteoporotic. It is a compound that researchers have become very interested in recently.

In our laboratory, we integrate molecular template, spray fluidized bed and electrostatic precipitation technology to create a novel spray fluidized bed polymerization method. This technique is to simplify processes in conventional molecular imprinted polymerization method. The concept is mixing target molecule, functional monomer, crosslinker, initiator and solvent and operating atomizer to produce aerosol. The suspension of the aerosol fluized polymerized by UV light in the quartz tube and collected on the plate by static electricity. The advantages of this method are to get independent genistein imprinted micro-nano sphere and to complete polymerization reaction efficiency.

In this study, genistein was chosen as template to prepare the molecularly imprinted polymer. The batch re-binding study and isothermal titration method were introduced to investigate the optimized formulation of genistein-imprinted polymer. Comparing the results of isothermal titration and batch re-binding study, TRIM and MAA were the optimized cross-linker and functional monomer among the monomers tested in this study. The best volume ratio of MAA to TRIM is 1:8. After the polymerization, extraction and rebinding steps, the genistein molecular imprinted polymer appeared to have absorption quantity 506.59μg/g, imprinting factor 1.738. From review paper, the range of particle size is from several to hundred micrometer during fluidization. In this research, the average size of MIP can reach 90nm during spray fluidized process from ZATASIZER results. The results showed that the selectivity of genistein-imprinted polymer was 50.5 in multi-component. In the soybean milk extraction rebinding test showed that the absorption genistein quantity can reach 80.92μg/g and absorption daidzein quantity can reach 39.77μg/g. This result revealed thst the imprinted polymer exhibit selectivity and affinity for genistein.
中文摘要…………………………………………………………………I
英文摘要………………………………………………………………III
誌謝………………………………………………………………………V
目錄………………………………………………………………………VI
表目錄…………………………………………………………………XII
圖目錄…………………………………………………………………XIII
專有名詞縮寫對照表………………………………………………XVIII
符號說明………………………………………………………………XIX
第一章 緒論………………………………………………………………1
1-1 豆漿之介紹…………………………………………………………1
1-2 異黃酮素……………………………………………………………3
1-2-1 異黃酮素之結構…………………………………………………3
1-2-2 異黃酮素之分析方法……………………………………………5
1-2-3 異黃酮素之萃取方法……………………………………………7
1-3 金雀異黃酮…………………………………………………………9
1-3-1 金雀異黃酮之來源………………………………………………9
1-3-2 金雀異黃酮之生理療效…………………………………………9
1-3-3 金雀異黃酮之生理代謝…………………………………………12
1-4 研究動機……………………………………………………………13
第二章 原理……………………………………………………………14
2-1 分子模版……………………………………………………………14
2-1-1 分子模版之起源與發展…………………………………………14
2-1-2 分之模版之原理…………………………………………………16
2-1-3 分子模版之材料…………………………………………………22
2-1-3.1 功能性單體之決定……………………………………………22
2-1-3.2 交聯劑之決定…………………………………………………24
2-1-3.3 溶劑之決定……………………………………………………26
2-1-3.4 起始劑之決定…………………………………………………27
2-1-4 分之模版之應用…………………………………………………28
2-1-5 分子模版記憶性吸附之動力式推導……………………………32
2-2 儀器原理……………………………………………………………34
2-2-1 高效能液相層析儀 (HPLC)……………………………………34
2-2-2 微熱卡計 (Isothermal titration calorimeter)…………35
2-2-3 掃描式電化學顯微鏡(Scanning Electron Microscope)……40
2-2-4 穿透式電子顯微鏡 (Transmission electron microscope, TEM)………42
2-2-5 雷光散射法粒徑分析儀 (zetasizer)…………………………44
2-2-6 比表面積測量原理 (BET)………………………………………45
2-2-7 傅立葉式紅外線吸收光譜儀……………………………………46
2-3 氣懸膠介紹…………………………………………………………47
2-3-1 氣懸膠之定義……………………………………………………47
2-3-2 氣懸膠之產生……………………………………………………47
2-3-3 氣懸膠之帶電行為………………………………………………48
2-4 靜電集塵原理………………………………………………………50
2-4-1 微粒帶電機制……………………………………………………50
2-4-2 微粒收集粒徑大小與裝置關係…………………………………52
第三章 實驗設備與方法………………………………………………54
3-1 藥品與儀器…………………………………………………………54
3-1-1 藥品………………………………………………………………54
3-1-2 儀器………………………………………………………………56
3-2 實驗流程與方法……………………………………………………57
3-2-1 噴霧流化床聚合系統……………………………………………57
3-2-1.1 聚合液之配製…………………………………………………57
3-2-1.2 噴霧流化床聚合流程…………………………………………57
3-2-1.3 金雀異黃酮的移除……………………………………………60
3-2-2 分子模版之再吸附測試…………………………………………60
3-2-2.1 校正曲線之製作………………………………………………60
3-2-2.2 單一成份系統測試……………………………………………60
3-2-2.3 多成份系統測試………………………………………………61
3-2-3高效能液相層析儀(HPLC)之操作………………………………61
3-2-4 微熱卡計(ITC)之等溫滴定……………………………………63
3-2-5 分子模版之特性分析……………………………………………65
3-2-5.1 掃描式電子顯微鏡分析………………………………………65
3-2-5.2 穿透式電子顯微鏡分析………………………………………65
3-2-5.3 比表面積分析…………………………………………………65
3-2-5.4 雷光散射法粒徑分析…………………………………………66
3-2-5.5 傅立葉轉換式紅外光譜儀……………………………………66
3-3 實驗架構與流程……………………………………………………67
第四章 實驗結果與討論………………………………………………68
4-1待測物校正曲線的製作……………………………………………68
4-1-1 金雀異黃酮之校正曲線…………………………………………70
4-1-2 大豆黃酮之校正曲線……………………………………………71
4-1-3 黃酮素之校正曲線………………………………………………72
4-2製備金雀異黃酮分子模版…………………………………………73
4-2-1利用微熱卡計找出最適之交聯劑………………………………73
4-2-2利用微熱卡計找出最適之功能性單體…………………………79
4-3各變因對製備金雀異黃酮分子模版之影響………………………84
4-3-1.1溶劑使用量對模版粒徑之影響………………………………84
4-3-1.2溶劑使用量對模版效能之影響………………………………96
4-3-2聚合方式對模版效能之影響……………………………………100
4-3-3-聚合所使用溶劑對模版效能之影響…………………………105
4-3-4吸附環境對模版效能之影響……………………………………110
4-4異黃酮分子之再吸附測試…………………………………………112
4-4-1異黃酮分子模版對干擾物之吸附測試…………………………112
4-4-1.1單成份系統下金雀異黃酮分子模版對干擾物之吸附………113
4-4-1.2多成份系統下金雀異黃酮分子模版對干擾物之吸附………115
4-5金雀異黃酮分子模版應用於豆漿中之吸附………………………117
4-5-1 豆漿萃取液內異黃酮素組成分析……………………………117
4-5-2分子模版對豆漿溶液之吸附效能評估…………………………120
第五章 综合討論………………………………………………………124
第六章 結論與建議……………………………………………………129
參考文獻………………………………………………………………131
附錄A 分子模版穿透式電子顯微鏡圖………………………………142
附錄B 傅立葉轉換式紅外光譜儀(FTIR)……………………………145
附錄C HPLC層析圖譜…………………………………………………146
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