蓖麻油基微乳劑作為蝦青素載體的配方和測試以及其用於外用給藥之可行性

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研究生：	吳勇美 Chintya Gunarto
論文名稱：	蓖麻油基微乳劑作為蝦青素載體的配方和測試以及其用於外用給藥之可行性 Formulation and testing of castor oil-based microemulsions as carrier for astaxanthin for possible application in topical drug delivery
指導教授：	朱義旭 Yi-Hsu Ju 吳耀豐 Alchris Woo Go
口試委員:	李振綱 Cheng-Kang Lee Felycia Edi Soetaredjo Suryadi Ismadji 翁玉鑽 Artik Elisa Angkawijaya 吳耀豐 Alchris Woo Go 朱義旭 Yi-Hsu Ju
學位類別：	博士 Doctor
系所名稱：	工程學院 - 化學工程系 Department of Chemical Engineering
論文出版年：	2021
畢業學年度：	109
語文別：	英文
論文頁數：	128
中文關鍵詞：	微乳液、蓖麻油、纖維素納米晶體、蝦青素、穩定、生物活性、釋放分布
外文關鍵詞：	microemulsion, castor oil, cellulose nanocrystals, astaxanthin, stability, biological activities, release profile
相關次數：	點閱：221 下載：2
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微乳液 (ME) 是一種由油和水組成，並添加了表面活性劑和助表面活性劑的膠體懸浮液。ME被探討其在各種食品、化妝品和藥物配方中的應用之可能。在本研究中以具有抗炎和鎮痛活性之蓖麻油為油相、Tween 80 或Tween 20為表面活性劑、甘油或乙醇為輔助表面活性劑，以及水相來形成ME。在表面活性劑混合物為Tween 80及乙醇；其重量比為2下可獲得的最大 ME 區域。 ME 的水力動力學直徑取決於油相和載藥量，纖維素奈米晶(CNC) 有潛力提高具有高蓖麻油含量的 ME 的穩定性。在自由基清除活性測試中，當 ME 配方中蓖麻油的比例從5 wt.%增加到28 wt.%時，IC50 的濃度從 494.47 µL/mL 降低到 52.25 µL/mL。在抗微生物試驗中觀察到相同的現象，分別可以抑制 94.9% 和 93.9% 的大腸桿菌和金黃色葡萄球菌的生長。從弗朗茲擴散法所得的12 小時蝦青素累積釋放量，在pH 為 7.4 和 5.6時分別為 11.17% 及 37.48%。蓖麻油含量為 10 和 15 wt.%在pH 5.6 的介質可得高百分比釋放，且使用 Sigmoidal 模型最佳描述此釋放曲線。 ME 配方中不同的初始加載量和 CNC 添加量會產生不同的釋放常數 (k) 和 ASX 釋放動力學的最大理論釋放 (Rmax)。以蓖麻油為基的 ME 對 HeLa 細胞的體外細胞毒性的初步研究顯示，在 200 µg/mL 和 7500 µg/mL 的濃度下，存活率分別僅為 76.36% 至 86.25% 和 27.44% 至 36.71%。這些結果證實了蓖麻油基微乳劑有可能作為蝦青素載體用於外部給藥，也可能用於治療癌細胞。

Microemulsion (ME) is a colloidal suspension that consists of oil and water, with addition of a surfactant and cosurfactant. It is explored for their use in various food, cosmetics, and drug formulations. In this study, castor oil (CO), which has anti-inflammatory and analgesic activities is used as the oil phase along with Tween 80 or Tween 20 as the surfactant, glycerol or ethanol as the cosurfactant, and aqueous phase in ME formulations. The largest ME region was attained at a surfactant mixture of Tween 80 and ethanol with weight ratio = 2. The hydrodynamic diameter of ME is dependent on the oil phase and amount of drug loaded, with cellulose nanocrystal (CNC) potentially improving the stability of ME having high CO fraction. The concentration at IC50 was reduced from 494.47 µL/mL to 52.25 µL/mL as the fraction of castor oil in ME formulation increased between 5~28 wt.% for free-radical scavenging activity. The same phenomenon was observed for anti-microbial assay, allowing 94.9 % and 93.9 % inhibition of E.coli and S.aureus growth, respectively. Cumulative ASX release obtained from Franz diffusion technique at 12 h ranged from 11.17 % to 37.48 % for pH 7.4 and 5.6, respectively. Castor oil with 10 and 15 wt.% resulted in high percentage release at pH medium of 5.6 and the release profile was best describe using a Sigmoidal model. Different initial loadings and addition of CNC in ME formulations gave different release constants (k) and maximum theoretical release (Rmax) of ASX release kinetic. Preliminary investigation on the in-vitro cytotoxicity of CO-based MEs on HeLa cells resulted in viabilities of only 76.36 % to 86.25 % and 27.44 % to 36.71 % at concentration of 200 µg/mL and 7500 µg/mL, respectively. These results confirmed that castor oil-based microemulsions could potentially be used as astaxanthin carrier for topical drug delivery and may also be used possibly be used in treatment of cancer cells.

摘要    ii
ABSTRACT    iii
ACKNOWLEDGEMENT    iv
TABLE OF CONTENTS    v
LIST OF ABBREVIATIONS    viii
LIST OF FIGURES    x
LIST OF TABLES    xiii
CHAPTER 1    1
1.    Background of the Study    1
2.    Goal and Objectives    3
3.    Significance of the Study    3
4.    Scope and Limitations    4
CHAPTER 2    5
1. Microemulsion    5
2. Components of a microemulsion    10
2.1. Oil    10
2.2. Surfactant    11
2.3. Cosurfactant    13
2.4. Pickering agent    14
3. Characteristics of microemulsion    15
3.1. Pseudoternary area    15
3.2. Hydrodynamic diameter    16
3.3. Storage stability    18
3.4. Biological activities    21
4. Hydrophobic drug: Astaxanthin    23
5. Drug or active compound release    25
5.1. Franz diffusion cell    26
5.2. Models to describe drug release profile    27
CHAPTER 3    30
1. Materials    30
2. Construction of pseudoternary phase diagram    32
3. Preparation of microemulsions    32
4. Physicochemical characteristics    32
5. Storage stability test    33
5.1. Heating-cooling stability test    33
5.2. Freeze-thaw stability test    34
5.3. Centrifugal stability test    34
6. Preparation of MEs-modified    34
6.1. Preparation of cellulose nanocrystal    34
6.2. Formulation of ASX loaded microemulsion    35
7. Biological activity assays    35
7.1. Free-radical scavenging activity    35
7.2. Anti-microbial activity    36
7.3. In-Vitro Cytotoxicity    36
8. Statistical and regression analysis    37
9. Drug release    38
9.1. Release study by Franz diffusion cell    38
9.2. Stability of ASX loaded-microemulsion    39
CHAPTER 4    41
1. Pseudoternary phase diagram    41
2. Characterization of ME    45
3. Astaxanthin loaded-microemulsion    48
4. Stability study of ME and ME@ASX    49
CHAPTER 5    53
1. Effect of HD on different formulations    53
2. Effect of astaxanthin loading on castor oil-based microemulsions    57
3. Effects of microemulsion formulation on the hydrodynamic diameter    57
4. Free-radical scavenging activity of castor oil-based microemulsions    61
5. Anti-microbial activity of castor oil-based microemulsions    62
6. Stability of castor oil-based microemulsions    64
CHAPTER 6    71
1. Stability of ASX loaded in castor-oil based microemulsions    71
2. Release or diffusion of ASX from ASX-loaded CO-based ME at different pH    76
3. Influence of loading on the release or diffusion of ASX from ASX-loaded CO-based MEs    80
4. Influence of CNC addition on release study    82
5. Cytotoxicity of castor oil-based microemulsions    83
CHAPTER 7    85
1. Conclusions    85
2. Recommendations    86
REFERENCES    87
APPENDIX    103


                                

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