臺灣博碩士論文加值系統

摘 要

本研究探討離子對雙親分子HTMA-DS、雙十六碳鏈磷酸鹽(DHDP)及膽固醇形成陰陽離子液胞的行為。陰陽離子液胞是以機械分散法生成，並評估做為穩定劑之DHDP和膽固醇的添加對液胞穩定性與包覆效率的影響。在此研究中，利用液胞雙層膜包覆親油性藥物維他命E，以了解與藥物/化妝品傳輸載體系統之配方相關的重要參數。此研究是以高效液相層析儀偵測維他命E的濃度，並使用螢光偏極化法探討液胞雙層膜的排列特性。
液胞之物理穩定性是在室溫下由肉眼觀察、液胞大小及界面電位的隨時間觀測來決定，發現此陰陽離子液胞系統可穩定長達14個月，液胞的平均大小在55到125奈米之間，且具窄的粒徑分布，由界面電位值-40到-63 mV可確認液胞帶負電性。膽固醇與雙親分子之頭基及疏水性碳鏈的交互作用，顯然在陰陽離子液胞的穩定形成上扮演重要的角色。帶負電之DHDP分子的添加，可透過液胞間的電性排斥力提供障礙而避免液胞的聚集，以促進液胞的物理穩定性。電子穿透顯微鏡影像顯示液胞呈球狀且大小均勻。根據液胞之包覆效率的高低，依序為添加43 mol%、50 mol%及35 mol%膽固醇的液胞，隨著DHDP莫爾分率的增加，對維他命E的包覆效率則隨之下降。膽固醇和DHDP的加入明顯影響了液胞的雙層膜性質及包覆效率。毒性測試則顯示隨著液胞中DHDP莫爾分率的增加，似會使陰陽離子液胞系統較不具毒性。

ABSTRACT

The formation characteristics of catanionic vesicles formed by ion pair amphiphile HTMA-DS, dihexadecyl phosphate (DHDP), and cholesterol were investigated in this study. Catanionic vesicles were prepared with a mechanical dispersion approach. The addition of DHDP and cholesterol as stabilizing agents was applied in order to evaluate their effects on the vesicle stability and encapsulation efficiency. In the present study, vitamin E as a lipophilic drug was encapsulated within the vesicular bilayers to elucidate the key parameters involved in the formulation of the drug/cosmetic carrier system. The concentration of vitamin E was measured by HPLC with UV detector and a fluorescence polarization study was carried out to investigate the bilayer packing characteristic.
Physical stability of the catanionic vesicles determined by visual observation, vesicle size, and vesicle zeta potential was monitored with time at room temperature. This catanionic vesicle system was found to be stable for more than fourteen months. The mean sizes of vesicles ranged from 55 nm to 125 nm with narrow size distributions. Negatively charged characteristic of the vesicles was confirmed based on the zeta potential values that varied from -40 mV to -63 mV. The interactions of cholesterol with the headgroups and hydrophobic chains of amphiphiles apparently played important roles in the formation of stable catanionic vesicles. Negative charges of DHDP molecules provided a barrier to prevent aggregation of the vesicles and increased their physical stability with the electrostatic repulsive force between the vesicles. Transmission electron microscopy (TEM) images showed the spherical structure and homogenous size of the vesicles. The encapsulation efficiency values of the vesicles were found to be in the order with the added cholesterol concentration of 43 mol%, 50 mol%, and 35 mol%. It has been observed that with a higher molar fraction of DHDP, the vitamin E encapsulation efficiency became lower. The incorporation of cholesterol and DHDP significantly affected the bilayer properties of the catanionic vesicles as well as the encapsulation efficiency. With the cytotoxicity experiment, it was suggested that incorporation of more DHDP into the catanionic vesicles resulted in the less toxicity of the catanionic vesicle systems.

Table of Contents
ABSTRACT i
摘 要 iii
ACKNOWLEDGEMENT iv
Table of Contents vi
List of Tables ix
List of Figures x
CHAPTER 1 INTRODUCTION 1
CHAPTER 2 LITERATURE REVIEW 4
2.1 Ion pair amphiphile 4
2.2 Charged vesicles 7
2.3 Effects of cholesterol 9
2.4 Encapsulation of vitamin E 12
CHAPTER 3 EXPERIMENTAL 19
3.1 Materials 19
3.2 Equipments 19
3.2.1 Ultrasonic dispersion device 19
3.2.2 Dynamic light scattering particle size analyzer 20
3.2.3 Laser doppler electrophoresis zeta potential analyzer 21
3.2.4 Transmission electron microscope 22
3.2.5 High performance liquid chromatography instrument 23
3.2.6 Multi-mode microplate reader 24
3.3 Methodology 24
3.3.1 Preparation of HTMA-DS 24
3.3.2 Preparation of catanionic vesicles 25
3.3.3 Size distribution of catanionic vesicles 26
3.3.4 Zeta potential of catanionic vesicles 26
3.3.5 Transmission electron microscope observation 26
3.3.6 Evaluation of vitamin E encapsulation efficiency 27
3.3.7 Fluorescence polarization measurement 28
3.3.8 Cell viability assay 29
CHAPTER 4 RESULTS AND DISCUSSION 34
4.1 Formation of catanionic vesicles 34
4.1.1 Catanionic vesicles formed by HTMA-DS 35
4.1.2 Catanionic vesicles formed by HTMA-DS/cholesterol 37
4.1.3 Catanionic vesicles formed by HTMA-DS/DHDP 38
4.1.4 Catanionic vesicles formed by HTMA-DS/DHDP/cholesterol 40
4.2 Encapsulation of vitamin E with catanionic vesicles 48
4.2.1 Properties of catanionic vesicles formed by HTMA-DS/DHDP/ cholesterol/vitamin E 49
4.2.2 Separation of vitamin E-loaded vesicles and non-encapsulated vitamin E 52
4.2.3 Encapsulation efficiency for vitamin E 53
4.3 Toxicity of catanionic vesicles formed by HTMA-DS/DHDP/cholesterol 59
CHAPTER 5 CONCLUSIONS 84
Appendix 87
REFERENCES 89
CURRICULUM VITAE 100

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