臺灣博碩士論文加值系統

微脂粒是在1960年代首先由英國科學家班韓姆（A. Bangham）所發現的，它是一種由脂雙層所構成類似細胞膜的中空球體，可將我們所感興趣的不同成份包覆其中。由於其特殊的性質和構造，使得微脂粒廣泛應用於生物學、製藥學以及醫學等方面的研究上。
本研究的目的在於製備食品級的微脂粒，希望以分子量相近的脂肪酸如stearic acid和linoleic acid以及分子構型較大的tristearin來取代傳統配方中的膽固醇，以減低口服微脂粒時膽固醇的攝取量，並探討經不同配方改良後的微脂粒，與傳統配方在包覆率、長效性、對pH的安定性及對酵素安定性等方面的影響。
結果發現，以去水-復水法所製備出的微脂粒為多層微脂粒（Multilamellar vesicles,MLV）。在微脂粒包覆牛血清蛋白（bovine serum albumin,BSA）之包覆效率（encapsulation efficiency,EE）實驗中發現，在微脂粒中添加stearic acid組(EE:55%)與添加膽固醇組(51%)其包覆效率皆明顯優於未添加組(44%)。添加tristearin組則無論其添加量為何，其包覆率在統計上均與未添加組無顯著差異存在；而添加linoleic acid組，隨著添加量的增加，其包覆率反而下降。以微脂粒包覆葡萄糖水溶液時，添加膽固醇、stearic acid、tristearin組均可增加其包覆率，且在lecithin : stearic acid或tristearin為1 : 0.5時達到最高值。另外將微脂粒經多次凍乾-復水的處理過程後可有效增加其包覆效率。
將微脂粒置於不同pH值（pH 3~10）的緩衝溶液中，發現微脂粒存於pH 6時，其包覆物質有一最低的滲出率。四種不同配方中以添加膽固醇組對酸鹼度的耐受性最佳，添加stearic acid 組次之。
將微脂粒長期儲存於-20℃的環境下，發現其包覆物質滲出率比儲存於4℃時大，而在微脂粒中添加α-tocopherol可有效降低其TBA值及滲出率。四種配方的滲出率以添加tristearin組最大，而添加膽固醇組在各種儲存溫度下都最穩定。
將α-amylase經微脂粒包覆後，添加膽固醇組和添加stearic acid組均可有效保護內部所包覆的活性物質不受強酸（pH 2.8）和胃蛋白酶的傷害，而添加tristearin組則無保護效果。
綜合以上結果顯示，在四種微脂粒的實驗配方中，添加stearic acid組其包覆效率最佳、對氧化安定性最穩且長效安定性僅次於添加膽固醇組，因此stearic acid可用以取代膽固醇來製備食品級的微脂粒。

Liposomes were first found by English scientist Dr. A. Bangham in 1960’s. Their specifically hollow and spherical structures are made up with multilayer cell-membrane-like phosphobilayers and are capable of encapsulating materials of interests. Such specific characteristics of liposomes lead to their application researches in various fields such as biology, pharmaceutics and medical science.
In the present study, liposomes were prepared for food use with various ratios of lecithin to stearic acid, linoleic acid or tristearin, inplace of the currently used cholesterol, in order to avoiding the excessive intake of cholesterol, In addition, their physico-chemical properties such as encapsulation efficiency（EE, %）and stabilities against storage, pH and pepsin were investigated to optimize the composition.
Liposomes of multilamellar vesicles（MLVs）were successfully prepared with the dehydration-rehydration method, and the products prepared with the addition of stearic acid（EE : 55%）or cholesterol （51%）exhibited higher encapsulation efficiency of bovine serum albumin（BSA）than those prepared without addition of oil （44%）, while those prepared with the addition of tristearin were insignificantly different with control, regardless of the amount of tristearin added. However, EE of BSA in liposomes decreased with the increasing amount of linoleic acid added. The EE of glucose in liposomes increased with the increasing level of cholesterol, stearic acid or tristearin added in liposomes, until a lecithin to stearic acid or tristearin ratio of 1：0.5 is reached, and then decreased. Besides, increasing dehydration-rehydration cycle in preparing liposomes was beneficial in increasing the EE.
Stability of liposomes against pH（pH 3 — 10） was determined by computing the released percentage of the encapsulated BSA. Results showed that liposomes at pH 6 were most stable, regardless of their compositions. Liposomes prepared with cholesterol added displayed the highest pH stability, followed by those prepared with stearic acid added.
Storage temperature was also influential on the released percentage of the encapsulated material. Storage at -20C exhibited more serious damage to the liposomes than at 4C. Addition of α-tocopherol was found effective in reducing the released percentage of encapsulated material by decreasing the TBA of the liposomes. Liposomes containing tristearin showed the highest released percentage, while those containing cholesterol were found stable during the 60-day storage at various temperatures.
Liposomes containing cholesterol or stearic acid were effective in protecting encapsulated α-amylase against acid (pH 2.8) and pepsin, while those containing tristearin were ineffective.
Among the four components tested, liposomes prepared with stearic acid added exhibited the highest EE and oxidation stability. Therefore, replacement of cholesterol by stearic acid in preparing liposomes for food use is feasible and recommended

目 錄
頁碼
中文摘要 ……………………………………………………Ⅰ
英文摘要 ……………………………………………………Ⅲ
前言 …………………………………………………….. 1
第Ⅰ章、文獻整理 ………………………………………… 3
一、 微脂粒（liposome）簡介 ………………………… 3
（1） 微脂粒的結構組成 ………………………… 3
（2） 微脂粒的分類 ……………………………….. 6
（3） 微脂粒的製備 ………………………………. 9
（4） 膽固醇添加對微脂粒的影響 ………………. 14
（5） 微脂粒於食品工業上的應用 ………………. 16
二、 微脂粒的包覆率 …………………………………. 19
三、 微脂粒的穩定性 …………………………………. 21
（1） 物理穩定性 ………………………………… 24
（2） 化學穩定性 ………………………………… 26
四、 微脂粒的長效性 ……………………………….…. 32
五、 膽固醇對人體的影響 ……………………………. .33
第Ⅱ章、材料與方法 …………………………………..…… 35
一、 藥品與器材 ……………………………………….. 35
（1） 藥品 ………………………………………… 35
（2） 器材 ………………………………………… 36
二、 儀器 ………………………………………………...37
三、 實驗方法 …………………………………………...38
（1） 微脂粒的製備 ……………………………….38
（2） 微脂粒的包覆率 …………………………… 39
（3） 蛋白質的定量 ……………………………… 39
（4） 微脂粒經多次凍乾-復水處理後之包覆率…42
（5） 葡萄糖的定量 ……………………………… 42
（6） 微脂粒的安定性 …………………………… 42
（7） pH值對微脂粒穩定性之影響 …………… 43
（8） 微脂粒的氧化 ……………………………….43
（9） 胃蛋白酶水解測定 ………………………….44
（10）α- amylase酵素活性分析 …………………44
（11）統計分析 …………………………………47
第Ⅲ章、結果與討論 ……………………………………….. 48
一、 微脂粒的包覆率 ………………………………….. 48
二、 微脂粒對不同pH值之耐受性 ……………………55
三、 微脂粒的粒徑分布 ………………………………...58
四、 微脂粒的長效性 …………………………………...64
五、 α-amylase經微脂粒包覆後對胃蛋白酶之耐受性 74
第Ⅳ章、結論 …………………………………………………78
第Ⅴ章、參考文獻 ……………………………………………80

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