本研究利用沉澱法，將 十 烷 基三甲基溴化銨(decyltrimethylammonium bromide, DeTMAB)和十四烷基硫酸鈉(sodium n-tetradecylsulfate, STS)以等莫爾混合，得到主要材料，離 子 對 雙 親 分 子 DeTMA-DS ( decyltrimethylammoniumdodecylsulfate )，接著用半自發製成，以乙醇當作共溶劑，並使用 pH7.4 的緩衝溶液(tris buffer)當作外在環境，以模擬人體血液，並添加膽固醇當作膜內添加劑，在不同比例的乙醇和膽固醇
濃度下製成液胞，並探討液胞包覆水溶性藥物熊果素(Arbutin)和油溶性藥物維他命 E 醋酸脂(dl-α-tocopheryl acetate, VEA)的物理特性，以及同時包覆兩種藥物和包覆單一種藥物的差異。實驗結果顯示，隨著膽固醇的添加，單一包覆熊果素液胞之粒徑與包覆效率有高度相關性，而單一包覆 VEA 液胞之雙層膜堅硬度與包覆效率有高度相關性，在這個基礎下，進行同時包覆實驗，並量測熊果素包覆效率與 VEA 包覆效率，結果發現，同時包覆熊果素與 VEA，且膽固醇莫耳分率在 0.3 以上時，其熊果素包覆效率都會比單一包覆熊果素來得高，然而，同時包覆熊果素與VEA，其 VEA 包覆效率與單一包覆 VEA 卻是差不多的。另外也發現當雙層膜中同時存在 VEA 與膽固醇時，雙層膜會比沒有包
覆 VEA 還要軟，即便同時包覆熊果素也是如此，因此，同時包覆同果素與 VEA 在經皮藥物傳輸方面非常有應用價值。

A simple semi-spontaneous process has been developed for fabricating ethosome-like catanionic vesicles for the transdermal delivery of drugs by using catanionic surfactant, a relatively high concentration of ethanol, and cholesterol (CHOL) in buffer solution. In this work, the vesicles were prepared with different ratios of ethanol and cholesterol concentration, and the physical properties and the encapsulation efficiency of water-soluble drug (arbutin) and oil soluble drug (vitamin E acetate) were discussed, then the difference between simultaneous and
individual encapsulation will be compared. The experimental results show that the arbutin encapsulation efficiency is highly correlated with the particle size, while the VEA encapsulation efficiency is highly correlated with the membrane rigidity. When arbutin and VEA are encapsulated simultaneously, it is found that when the molar fraction of cholesterol is above 0.3, the arbutin encapsulation efficiency will be higher than that of individual encapsulation of arbutin. However, the VEA encapsulation efficiency is almost the same between simultaneous and individual
encapsulation. In addition, it was found that when the molar fraction of cholesterol is above 0.3, There is the interaction between VEA and cholesterol in the bilayer, so the membrane rigidity will be smaller. The results achieved in this work is that simultaneous encapsulation is not only more convenient for application, but also benefit to the encapsulation efficiency of arbutin, so it has great potential for transdermal drug delivery.

1. Jesorka, A. and O. Orwar, Liposomes: technologies and analytical applications. Annu. Rev. Anal. Chem., 2008. 1: p. 801-832.
2. Ahmad, I., et al., Antibody-targeted delivery of doxorubicin entrapped in sterically stabilized liposomes can eradicate lung cancer in mice. Cancer research, 1993. 53(7): p. 1484-1488.
3. Karmali, P.P. and A. Chaudhuri, Cationic liposomes as non‐viral carriers of gene medicines: resolved issues, open questions, and future promises. Medicinal research reviews, 2007. 27(5): p. 696-722.
4. Deepthi, V. and A. Kavitha, Liposomal drug delivery system–a review. Rguhs J. Pharm. Sci, 2014. 4(2): p. 47-56.
5. Mouritsen, O.G., Lipids, curvature, and nano‐medicine. European journal of lipid science and technology, 2011. 113(10): p. 1174-1187.
6. Dhawan, V.V. and M.S. Nagarsenker, Catanionic systems in nanotherapeutics–Biophysical aspects and novel trends in drug delivery applications. Journal of Controlled Release, 2017. 266: p. 331-345.
7. Subedi, R.K., et al., Recent advances in transdermal drug delivery. Archives of Pharmacal Research, 2010. 33(3): p. 339-351.
8. Cevc, G., Lipid vesicles and other colloids as drug carriers on the skin. Advanced drug delivery reviews, 2004. 56(5): p. 675-711.
9. Kaler, E.W., et al., Spontaneous vesicle formation in aqueous mixtures of single-tailed surfactants. Science, 1989. 245(4924): p. 1371-1374.
10. Koehler, R.D., S.R. Raghavan, and E.W. Kaler, Microstructure and dynamics of wormlike micellar solutions formed by mixing cationic and anionic surfactants. The Journal of Physical Chemistry B, 2000. 104(47): p. 11035-11044.
11. Lee, J.-H., et al., Vesicle− Biopolymer Gels: Networks of Surfactant Vesicles Connected by Associating Biopolymers. Langmuir, 2005. 21(1): p. 26-33.
12. Marques, E., et al., Self-organization of double-chained and pseudodouble-chained surfactants: counterion and geometry effects. Advances in colloid and interface science, 2003. 100: p. 83-104.
13. Chien, C., et al., Formation and encapsulation of catanionic vesicles. J. Chin. Colloid Interface Soc, 2002. 24: p. 31-45.
14. Yeh, S.-J., Y.-M. Yang, and C.-H. Chang, Cosolvent effects on the stability of catanionic vesicles formed from ion-pair amphiphiles. Langmuir, 2005. 21(14): p. 6179-6184.
15. Lee, W.-H., et al., Synthesis of ion-pair amphiphiles and calorimetric study on the gel to liquid-crystalline phase transition behavior of their bilayers. Journal of Chemical & Engineering Data, 2015. 60(4): p. 1119-1125.
16. Wu, K.-C., et al., Enhancement of catansome formation by means of cosolvent effect: Semi-spontaneous preparation method. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 2007. 302(1-3): p. 599-607.
17. Yang, Y.-M., et al., On the stability of liposomes and catansomes in aqueous alcohol solutions. Langmuir, 2008. 24(5): p. 1695-1700.
18. Israelachvili, J.N., Intermolecular and surface forces. 2011: Academic press.
19. Huang, J.-B. and G.-X. Zhao, Formation and coexistence of the micelles and vesicles in mixed solution of cationic and anionic surfactant. Colloid and Polymer Science, 1995. 273(2): p. 156-164.
20. Wang, C., et al., Transformation from precipitates to vesicles in mixed cationic and anionic surfactant systems. Colloid and Polymer Science, 2002. 280(8): p. 770-774.
21. Huang, J.-B., et al., Vesicle formation of a 1: 1 catanionic surfactant mixture in ethanol solution. Langmuir, 1997. 13(21): p. 5759-5761.
22. Huang, J.-B., et al., Vesicle formation of 1: 1 cationic and anionic surfactant mixtures in nonaqueous polar solvents. Colloid and Polymer Science, 1999. 277(4): p. 354-360.
23. Zhang, X.-R., et al., From precipitation to vesicles: a study on self-organized assemblies by alkylammonium and its mixtures in polar solvents. Colloid and Polymer Science, 2001. 279(12): p. 1245-1249.
24. Yu, W.-Y., Y.-M. Yang, and C.-H. Chang, Cosolvent effects on the spontaneous formation of vesicles from 1: 1 anionic and cationic surfactant mixtures. Langmuir, 2005. 21(14): p. 6185-6193.
25. Lasic, D.D., The mechanism of vesicle formation. Biochemical Journal, 1988. 256(1): p. 1-11.
26. Barry, J.A. and K. Gawrisch, Direct NMR evidence for ethanol binding to the lipid-water interface of phospholipid bilayers. Biochemistry, 1994. 33(26): p. 8082-8088.
27. Touitou, E., et al., Ethosomes—novel vesicular carriers for enhanced delivery: characterization and skin penetration properties. Journal of controlled release, 2000. 65(3): p. 403-418.
28. Huang, Z.-L., et al., Gelation of charged catanionic vesicles prepared by a semispontaneous process. Langmuir, 2010. 26(4): p. 2374-2382.
29. Liu, Y.-S., C.-F. Wen, and Y.-M. Yang, Cholesterol effects on the vesicular membrane rigidity and drug encapsulation efficiency of ethosome-like catanionic vesicles. Science of Advanced Materials, 2014. 6(5): p. 954-962.
30. 邱文昱, 陰陽離子液胞包覆油/水溶性藥物之行為探討. 成功大學化學工程學系學位論文, 2012: p. 1-132.
31. 温智芳, 經皮藥物傳輸用類乙醇體陰陽離子液胞之研發. 成功大學化學工程學系學位論文, 2013: p. 1-104.
32. 葉如萍, 類乙醇體陰陽離子液胞的雙層膜特性與其包覆/釋放行為的關聯. 成功大學化學工程學系學位論文, 2013: p. 1-121.
33. 邱俊瑋, 類乙醇體陰陽離子液胞的包覆/釋放與膠化行為之探討. 成功大學化學工程學系學位論文, 2013: p. 1-170.
34. 張詰苡, 以螢光非等向性方法檢視類乙醇體陰陽離子液胞的雙層膜堅硬度-乙醇及膽固醇效應探討. 成功大學化學工程學系學位論文, 2019: p. 1-92.
35. 王俊為, 單一雙層陰陽離子液胞的水溶性藥物包覆效率之理論與實驗比較研究-乙醇及膽固醇效應. 成功大學化學工程學系學位論文, 2018: p. 1-94.
36. Petrache, H.I., et al., Salt screening and specific ion adsorption determine neutral-lipid membrane interactions. Proceedings of the National Academy of Sciences, 2006. 103(21): p. 7982-7987.
37. Evans, E. and D. Needham, Physical properties of surfactant bilayer membranes: thermal transitions, elasticity, rigidity, cohesion and colloidal interactions. Journal of Physical Chemistry, 1987. 91(16): p. 4219-4228.
38. Grasso, D., et al., A review of non-DLVO interactions in environmental colloidal systems. Reviews in Environmental Science and Biotechnology, 2002. 1(1): p. 17-38.
39. Sabın, J., et al., Size and stability of liposomes: a possible role of hydration and osmotic forces. The European Physical Journal E, 2006. 20(4): p. 401-408.
40. Brown, M.F., et al., Elastic deformation of membrane bilayers probed by deuterium NMR relaxation. Journal of the American Chemical Society, 2002. 124(28): p. 8471-8484.
41. Walz, J.Y. and E. Ruckenstein, Comparison of the van der Waals and undulation interactions between uncharged lipid bilayers. The Journal of Physical Chemistry B, 1999. 103(35): p. 7461-7468.
42. Mannock, D.A., R.N. Lewis, and R.N. McElhaney, Comparative calorimetric and spectroscopic studies of the effects of lanosterol and cholesterol on the thermotropic phase behavior and organization of dipalmitoylphosphatidylcholine bilayer membranes. Biophysical journal, 2006. 91(9): p. 3327-3340.
43. Blandamer, M.J., et al., Vesicle-cholesterol interactions: Effects of added cholesterol on gel-to-liquid crystal transitions in a phospholipid membrane and five dialkyl-based vesicles as monitored using DSC. Physical Chemistry Chemical Physics, 2003. 5(23): p. 5309-5312.
44. Stillwell, W., et al., Cholesterol versus α-tocopherol: Effects on properties of bilayers made from heteroacid phosphatidylcholines. Biochemistry, 1996. 35(41): p. 13353-13362.
45. Malcolmson, R., et al., DSC of DMPC liposomes containing low concentrations of cholesteryl esters or cholesterol. Journal of membrane science, 1997. 123(2): p. 243-253.
46. Benesch, M.G. and R.N. McElhaney, A comparative calorimetric study of the effects of cholesterol and the plant sterols campesterol and brassicasterol on the thermotropic phase behavior of dipalmitoylphosphatidylcholine bilayer membranes. Biochimica et Biophysica Acta (BBA)-Biomembranes, 2014. 1838(7): p. 1941-1949.
47. Benesch, M.G., R.N. Lewis, and R.N. McElhaney, A calorimetric and spectroscopic comparison of the effects of cholesterol and its immediate biosynthetic precursors 7-dehydrocholesterol and desmosterol on the thermotropic phase behavior and organization of dipalmitoylphosphatidylcholine bilayer membranes. Chemistry and physics of lipids, 2015. 191: p. 123-135.
48. Aramaki, K., et al., Charge boosting effect of cholesterol on cationic liposomes. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 2016. 506: p. 732-738.
49. Bhattacharya, S. and S. Haldar, Interactions between cholesterol and lipids in bilayer membranes. Role of lipid headgroup and hydrocarbon chain–backbone linkage. Biochimica et Biophysica Acta (BBA)-Biomembranes, 2000. 1467(1): p. 39-53.
50. 鄭嘉瑜, 以螢光偏極化技術探討膽固醇對離子對雙親分子液胞雙層膜堅硬度的影響. 成功大學化學工程學系學位論文, 2019: p. 1-111.
51. 謝佑翎, 乙醇與膽固醇對 DPPC 脂質體雙層膜特性之影響. 成功大學化學工程學系學位論文, 2017: p. 1-101.
52. Lönnfors, M., et al., Interaction of 3β-amino-5-cholestene with phospholipids in binary and ternary bilayer membranes. Langmuir, 2012. 28(1): p. 648-655.
53. Silva, C., et al., Molecular aspects of the interaction between plants sterols and DPPC bilayers: an experimental and theoretical approach. Journal of colloid and interface science, 2011. 358(1): p. 192-201.
54. Krause, M.R., S. Turkyilmaz, and S.L. Regen, Surface occupancy plays a major role in cholesterol’s condensing effect. Langmuir, 2013. 29(33): p. 10303-10306.
55. Bui, T.T., K. Suga, and H. Umakoshi, Roles of sterol derivatives in regulating the properties of phospholipid bilayer systems. Langmuir, 2016. 32(24): p. 6176-6184.
56. El Khoury, E. and D. Patra, Length of hydrocarbon chain influences location of curcumin in liposomes: Curcumin as a molecular probe to study ethanol induced interdigitation of liposomes. Journal of Photochemistry and Photobiology B: Biology, 2016. 158: p. 49-54.
57. McMullen, T.P., R.N. Lewis, and R.N. McElhaney, Differential scanning calorimetric and Fourier transform infrared spectroscopic studies of the effects of cholesterol on the thermotropic phase behavior and organization of a homologous series of linear saturated phosphatidylserine bilayer membranes. Biophysical journal, 2000. 79(4): p. 2056-2065.
58. McMullen, T.P., R.N. Lewis, and R.N. McElhaney, Calorimetric and spectroscopic studies of the effects of cholesterol on the thermotropic phase behavior and organization of a homologous series of linear saturated phosphatidylglycerol bilayer membranes. Biochimica et Biophysica Acta (BBA)-Biomembranes, 2009. 1788(2): p. 345-357.
59. Bhattacharya, S. and S. Haldar, The effects of cholesterol inclusion on the vesicular membranes of cationic lipids. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1996. 1283(1): p. 21-30.
60. Smondyrev, A.M. and M.L. Berkowitz, Molecular dynamics simulation of the structure of dimyristoylphosphatidylcholine bilayers with cholesterol, ergosterol, and lanosterol. Biophysical journal, 2001. 80(4): p. 1649-1658.
61. Tien, W.-j., et al., Effects of Cholesterol on Water Permittivity of Biomimetic Ion Pair Amphiphile Bilayers: Interplay between Membrane Bending and Molecular Packing. International journal of molecular sciences, 2019. 20(13): p. 3252.
62. Yang, J., J. Martí, and C. Calero, Pair interactions among ternary DPPC/POPC/cholesterol mixtures in liquid-ordered and liquid-disordered phases. Soft Matter, 2016. 12(20): p. 4557-4561.
63. de Meyer, F. and B. Smit, Effect of cholesterol on the structure of a phospholipid bilayer. Proceedings of the National Academy of Sciences, 2009. 106(10): p. 3654-3658.
64. Xiang, T.-X. and B.D. Anderson, Phase structures of binary lipid bilayers as revealed by permeability of small molecules. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1998. 1370(1): p. 64-76.
65. 吳旌豪, 以螢光偏極化技術探討乙醇及膽固醇對陰陽離子液胞雙層膜堅硬度的影響. 成功大學化學工程學系學位論文, 2020: p. 1-117.
66. Kranenburg, M., M. Vlaar, and B. Smit, Simulating induced interdigitation in membranes. Biophysical journal, 2004. 87(3): p. 1596-1605.
67. Michl, J. and E.W. Thulstrup, Spectroscopy with polarized light: solute alignment by photoselection, liquid crystal, polymers, and membranes. Corrected Software Edition. Spectroscopy, 1995.
68. Masters, B.R., Molecular fluorescence: principles and applications. Journal of Biomedical Optics, 2013. 18(3): p. 039901.
69. Vincent, M., et al., Nanosecond fluorescence anisotropy decays of n-(9-anthroyloxy) fatty acids in dipalmitoylphosphatidylcholine vesicles with regard to isotropic solvents. Biochemistry, 1982. 21(4): p. 708-716.
70. Lakowicz, J.R., Principles of fluorescence spectroscopy. 2013: Springer science & business media.
71. Aung, N.N., et al., HPMC/PVP dissolving microneedles: a promising delivery platform to promote trans-epidermal delivery of alpha-arbutin for skin lightening. Aaps Pharmscitech, 2020. 21(1): p. 1-13.
72. Padamwar, M.N. and V.B. Pokharkar, Development of vitamin loaded topical liposomal formulation using factorial design approach: drug deposition and stability. International journal of pharmaceutics, 2006. 320(1-2): p. 37-44.
73. Gallarate, M., et al., Deformable Liposomes as Topical Formulations Containing α‐Tocopherol. Journal of dispersion science and technology, 2006. 27(5): p. 703-713.
74. Massey, J.B., Interfacial properties of phosphatidylcholine bilayers containing vitamin E derivatives. Chemistry and physics of lipids, 2001. 109(2): p. 157-174.
75. Chang, W.-H., et al., Effects of sterol-like additives on phase transition behavior of ion-pair amphiphile bilayers. Journal of oleo science, 2017. 66(11): p. 1229-1238.
76. Fukuda, H., et al., Bilayer-forming ion pair amphiphiles from single-chain surfactants. Journal of the American Chemical Society, 1990. 112(4): p. 1635-1637.
77. 柯政遠, 乙醇體及陰陽體的製備及其包覆/釋放行為之探討. 成功大學化學工程學系學位論文, 2008: p. 1-139.
78. Xu, X., M.A. Khan, and D.J. Burgess, Predicting hydrophilic drug encapsulation inside unilamellar liposomes. International journal of pharmaceutics, 2012. 423(2): p. 410-418.
79. Bhardwaj, U. and D.J. Burgess, Physicochemical properties of extruded and non-extruded liposomes containing the hydrophobic drug dexamethasone. International journal of pharmaceutics, 2010. 388(1-2): p. 181-189.
80. Duclairoir, C., et al., α-Tocopherol encapsulation and in vitro release from wheat gliadin nanoparticles. Journal of microencapsulation, 2002. 19(1): p. 53-60.
81. Liu, N. and H.-J. Park, Chitosan-coated nanoliposome as vitamin E carrier. Journal of microencapsulation, 2009. 26(3): p. 235-242.
82. Lopez-Pinto, J., M. Gonzalez-Rodriguez, and A. Rabasco, Effect of cholesterol and ethanol on dermal delivery from DPPC liposomes. International journal of pharmaceutics, 2005. 298(1): p. 1-12.
83. Ola, H., S.A. Yahiya, and O.N. El-Gazayerly, Effect of formulation design and freeze-drying on properties of fluconazole multilamellar liposomes. Saudi pharmaceutical journal, 2010. 18(4): p. 217-224.
84. Deniz, A., et al., Celecoxib-loaded liposomes: effect of cholesterol on encapsulation and in vitro release characteristics. Bioscience reports, 2010. 30(5): p. 365-373.
85. Briuglia, M.-L., et al., Influence of cholesterol on liposome stability and on in vitro drug release. Drug delivery and translational research, 2015. 5(3): p. 231-242.
86. Srivastava, S., et al., Fluidity, permeability and antioxidant behaviour of model membranes incorporated with α-tocopherol and vitamin E acetate. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1983. 734(2): p. 353-362.
87. Bradford, A., et al., The effect of vitamin E on the structure of membrane lipid assemblies. Journal of lipid research, 2003. 44(10): p. 1940-1945.
88. Gürsoy, A., E. Kut, and S. Özkırımlı, Co-encapsulation of isoniazid and rifampicin in liposomes and characterization of liposomes by derivative spectroscopy. International journal of pharmaceutics, 2004. 271(1-2): p. 115-123.
89. Ong, J.C.-L., F. Sun, and E. Chan, Development of stealth liposome coencapsulating doxorubicin and fluoxetine. Journal of liposome research, 2011. 21(4): p. 261-271.
90. Liu, J.J., et al., Preparation and characterization of cosmeceutical liposomes loaded with avobenzone and arbutin. Journal of cosmetic science, 2013. 64(1): p. 9-17.
91. Tsai, W.-C. and S.S. Rizvi, Simultaneous microencapsulation of hydrophilic and lipophilic bioactives in liposomes produced by an ecofriendly supercritical fluid process. Food research international, 2017. 99: p. 256-262.
92. Chen, W., et al., Co‐encapsulation of EGCG and quercetin in liposomes for optimum antioxidant activity. Journal of food science, 2019. 84(1): p. 111-120.
93. Huang, M., et al., Liposome co-encapsulation as a strategy for the delivery of curcumin and resveratrol. Food & function, 2019. 10(10): p. 6447-6458.
94. Hudiyanti, D., et al. Cholesterol implications on coconut liposomes encapsulation of beta-carotene and vitamin C. in IOP Conference Series: Materials Science and Engineering. 2019. IOP Publishing.
95. Nkanga, C.I., et al., Co-encapsulation of rifampicin and isoniazid in crude soybean lecithin liposomes. South African Journal of Chemistry, 2019. 72: p. 80-87.
96. Romero-Arrieta, M.R., E. Uria-Canseco, and S. Perez-Casas, Simultaneous encapsulation of hydrophilic and lipophilic molecules in liposomes of DSPC. Thermochimica Acta, 2020. 687: p. 178462.
97. Gent, M. and J. Prestegard, Cholesterol-phosphatidylcholine interactions in vesicle systems. Implication of vesicle size and proton magnetic resonance line-width changes. Biochemistry, 1974. 13(19): p. 4027-4033.
98. Subczynski, W.K., et al., Hydrophobic barriers of lipid bilayer membranes formed by reduction of water penetration by alkyl chain unsaturation and cholesterol. Biochemistry, 1994. 33(24): p. 7670-7681.
99. FUKAHORI, M., et al., Effects of added lower alcohols on solubilized tocopherol acetate sorption to polyethylene film in aqueous solution. Chemical and pharmaceutical bulletin, 1996. 44(1): p. 196-200.
100. Barry, J.A. and K. Gawrisch, Effects of ethanol on lipid bilayers containing cholesterol, gangliosides, and sphingomyelin. Biochemistry, 1995. 34(27): p. 8852-8860.
101. Lee, H., Vitamin E acetate as linactant in the pathophysiology of EVALI. Medical hypotheses, 2020. 144: p. 110182.