以改良式酒精注射法製備帶不同電性液胞包覆維生素E及其物化性質之研究

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標題:	以改良式酒精注射法製備帶不同電性液胞包覆維生素E及其物化性質之研究 Vesicles with different surface charges prepared by a modified ethanol injection method for encapsulation of vitamin E and their physicochemical properties
作者:	李昱頡
貢獻者:	嘉南藥理科技大學：化妝品科技研究所周宗翰
關鍵字:	改良式酒精注射法維生素E 不同帶電性 different surface charges modified ethanol injection method vitamin E
日期:	2011
上傳時間:	2011-10-25 13:27:57 (UTC+8)
摘要:	本研究主要以帶正電陽離子型界面活性劑，PM，PH及PO(統稱PX)之不同碳鏈長度混合帶負電NH，施以改良式酒精注射法形成液胞。使用動態光散射儀(dynamic light scattering, DLS)測量粒徑及界面電位，螢光分光光度計分析液胞疏水區間內及靠近親水頭基分子之流動性，和微分掃描熱卡計(differential scanning calorimetry, DSC)得出液胞從膠態轉為液晶態的相變溫度。再者，透過細胞存活度試驗(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT assay)以探討液胞對於人類皮膚纖維母細胞Hs68細胞的影響。並且觀察室溫下混合液胞的長時間儲存穩定天數。混合PX/NH液胞物化性質的結果顯示XPX＝0.9皆有較小的粒徑及XPX＝0.3、0.4則有較大的粒徑，液胞界面電位範圍從＋45 mV到－50 mV。液胞組成以XPM＝0、0.1、0.2、0.5、0.6、0.7、0.8、0.9，XPH＝0、0.1、0.2、0.4、0.5、0.6及XPO＝0、0.1、0.5、0.6、0.7、0.8、0.9、1較穩定，且穩定天數目前已達500天。陽離子型液胞PX 的膜內及頭基分子流動性皆較陰離子型液胞NH大，膜內流動性由高到低分別為PM>PH>PO>NH。純成分液胞主要相變溫度分別為PM＝29.7˚C，PH＝42.6˚C，PO＝52.9˚C，NH＝74.4˚C。細胞存活度結果顯示在濃度0.5 mM作用下，XPM＝0.3、0，XPH＝0.5、0.3、0.1、0，XPO＝0.7、0.5、0.3、0.1、0組成之液胞，細胞存活度在60%以上，純成分液胞毒性大到小依序為PM>PH>PO>NH。在應用方面，液胞包覆維生素E效率結果顯示XPM＝0.9及1， XPH＝0.7及0.9，XPO＝0.7及0.9其包覆維生素E可達30% (維生素E/界面活性劑)。純成分液胞包覆維生素E效率順序為PM>PH>PO>NH。此外，藉由Langmuir trough單分子層技術，探討液胞組成分子之間的交互作用力，並利用螢光顯微鏡觀察單分子層在氣/液界面上的型態排列變化。結果顯示陽離子界面活性劑的極限分子面積(limiting molecular area)較陰離子界面活性劑大，依序為PM>PH>PO>NH。在螢光顯微鏡影像中純成分PX單分子層呈現均勻黑色區塊，但純成分NH單分子層則呈現不規則白色雲霧狀。 In this study, cationic surfactants with different length, PX and NH, were used to prepare catanionic vesicles by a modified ethanol injection. This report used dynamic light scattering (DLS) to measure size, fluorescence polarization to investigate the membrane fluidity of vesicles inside and outside, and differential scanning calorimetry (DSC) to determine the gel to liquid-crystal phase transition of vesicles. And using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, to estimate the influence of catanionic vesicles for Hs68 cells. After the above examination of physicochemical characteristics, short-storage stability of catanionic vesicles was determined at room temperature. The results of physicochemical properties of mixed PX/NH vesicles showed XPX＝0.9 has a smaller particle size and XPX＝0.3 and 0.4 has a larger particle size, respectively. And the rang of zeta potential of vesicles from ＋45 mV to －50 mV. Vesicle with XPM＝0, 0.1, 0.2, 0.5, 0.6, 0.7, 0.8, 0.9, XPH＝0, 0.1, 0.2, 0.4, 0.5, 0.6 and XPO＝0, 0.1, 0.5, 0.6, 0.7, 0.8, 0.9 ,1 were stable and has been storaged over 500 days. Fluorescence polarization results shows the inside and outside membrane fluidity of PX was higher than that of NH, degree of pure vesicles membrane fluidity was PM>PH>PO>NH. Differential scanning calorimetry results showed the main phase transition temperature PM＝29.7˚C, PH＝42.6˚C, PO＝52.9˚C and NH＝74.4˚C. Cell viability showed at a concentration of 0.5mM, the viability of XPM＝0.3 and 0 XPH＝0.5, 0.3, 0.1, 0, and XPO＝0.7, 0.5, 0.3, 0.1, 0, were up to 60%, a decrease in cell viability of pure vesicles was PM>PH>PO>NH. In addition, these vesicles were applied to encapsulate vitamin E and the encapsulation efficiency was assayed by HPLC. The results shows that the XPM＝0.9 and 1, XPH＝0.7 and 0.9, and XPO＝0.7 and 0.9 has the high encapsulation efficiency of 30%. The entrapment efficiency of pure PX vesicles was PM>PH>PO>NH. A Langmuir trough monolayer technique was used to investigate intermolecular interaction between PX and NH, and combined with a microscope to observ monolayer brhavior at the air/water interface. The results shows limiting molecular area of PX was more than NH, and PM>PH>PO>NH. From fluorescence images, one can observed pure PX monolayers domains were dark and the morphology of pure NH monolayer was a clouds.
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