臺灣博碩士論文加值系統

前人研究藏紅花酸 (crocetin) 具有多種生物活性例如：增強氧擴散速率、抗氧化、抗發炎、抗高血壓、抗血栓、抗動脈粥狀硬化、抑制視網膜損傷和改善睡眠質量。雖然藏紅花酸具有許多生物活性，但藏紅花酸難溶於水造成口服藏紅花低的生物可利用率。因此開發適合藏紅花酸包埋的方法來提高生物可利用率。

根據張銘聰博士所建立的包埋薑黃素和油體固定化的技術將此加以改良成適合藏紅花酸包埋的方法。形成水包油的 (O/W) 奈米乳液包埋 (encapsulation) 藏紅花酸，先將藏紅花酸分散於 Span® 80親脂性介面活性劑中，加入芝麻油 (sesame oil) 和 IGEPAL® CO-720 親水性介面活性劑，最後加入磷酸鈉緩衝溶液 (10 mM) 和檸檬酸鈉溶液用超音波震盪混和彼此製成奈米乳液。為了測試埋率，將奈米乳液加在蔗糖溶液(40%)上方並高速離心，用高效液相層析方法分析糖水中未包埋的藏紅花酸濃度，換算後包埋率可達到99%。長時間久放造成奈米乳液的絮凝現象，因此將奈米乳液進行固定化 (solidfication)來避免聚集。固定化以加入不同比例甘露醇作為賦形劑並進行冷凍乾燥成粉末。實驗結果發現加入13.3%濃度的甘露醇固化後回溶的奈米乳液為最佳，並透過顯微鏡觀察發先固定化前後的奈米乳液仍可維持穩定分離的微胞達3個月，利用動態光散射儀量測的奈米乳液和回溶固定化奈米乳液的粒徑大小(平均粒徑: 124 ~ 343nm)。包埋率會因放置時間越長而下降，三個月後以 3.3% 甘露醇固定化組下降最多有 24%。
進行動物實驗以每組四隻大鼠分成對照組和實驗組，對照組餵食50 mg/kg 藏紅花酸的5%羧甲基纖維素鈉 (CMC-Na) 分散，實驗組餵食包埋後25 mg/kg藏紅花酸奈米乳液，管餵前先禁食12小時，取0、30、60、90、120、240、360分鐘的血液並以HPLC分析和作時間和血漿中藏紅花酸濃度圖。包埋藏紅花酸的奈米乳液吸收率比直接口服藏紅花酸好。

Previous studies have demonstrated that crocetin possesses various biological activities such as enhancing oxygen diffusion rate, antioxidant, anti-inflammatory, antihypertensive, antithrombotic, anti-atherosclerotic, inhibiting retinal damage and improve the quality of sleep. Although crocetin possesses several types of bioactivity, crocetin is poorly soluble in water and leads to low oral bioavailability. Therefore, development of crocetin via encapsulation enhances oral bioavailability of crocetin.
According as Dr. Ming-Tsung Chang established techniques of encapsulated curcumin and solidified oil bodies, so these were adjusted to be suitable for encapsulated crocetin . Forming oil-in-water (O/W) nanoemulsion encapsulates crocetin. Crocetin is dispersed in Span® 80 of lipophilic surfactant, then added with sesame oil and IGEPAL® CO-720 as hydrophilic surfactant. Finally, sodium phosphate buffer solution (10 mM) and sodium citrate solution are added and mixed to form nanoemulsion by ultrasonication. In order to test the encapsulation efficiency, the nanoemulsion was added on the sucrose solution (40%) with high-speed centrifugation. Analysis of non-encapsulated crocetin in sucrose solution was performed by HPLC method. The calculation of the encapsulation efficiency reaches to 99%. Prolonged storage causes flocculation of nanoemulsion, so nanoemulsion is solidified to avoid aggregation. Solidification was performed by adding different proportions of mannitol as excipients and lyophilisation as powder. The experiment discovered that 13.3% of mannitol redissolved well. Under microscopy examination, the solidfication of nanoemulsion maintained separation with micelle type until 3 months. The particle size of the nanoemulsion and the solidification nanoemulsion were measured by dynamic light scattering (average particle size range: 124 ~ 343 nm). The encapsulation efficiency is decreased in the long time storage .3.3% mannitol solidification decreased to 24% after three month.
Rats were divided into control and experimental groups. The control groups were fed crocetin 50 mg/kg and crocetin was dispersed in 5% CMC-Na (Sodium Carboxymethyl Cellulose) .The experimental groups were fed crocetin 25mg/kg of nanoemulsion. Before rats were fasted in 12 hours, rats were fed in gavage.0, 30, 60, 90, 120, 240, 360 minutes of blood samples were taken and analyzed by HPLC and made time and plasma concentrations of crocetin. The absorption of crocetin via nanoemulsion is better than directly oral crocetin.

壹.緒論…………1
貳.文獻回顧…………2
一、藏紅花苷(CROCIN)藏紅花酸(CROCETIN)來源和應用…………2
(一)梔子和水梔子…………2
(二)番紅花…………4
(三)物理化學性質…………6
(四)生物活性…………9
(五)藥物動力學相關研究…………11
二、脂溶性藥物吸收代謝途徑…………16
三、奈米乳液研究進展…………20
參.料與方法…………24
一、實驗設計與流程…………24
二、實驗材料…………25
(一)化學試劑…………25
(二)動物實驗及耗材…………26
(三)儀器設備…………26
三、實驗方法…………27
(一)藏紅花苷鹼水解得藏紅花酸…………27
(二)奈米乳液包埋藏紅花酸和固定化…………28
(三)粒徑測量和包埋率測試實驗…………30
(四)動物實驗…………31
肆.實驗結果與討論…………33
一、藏紅花苷鹼水解得藏紅花酸…………33
二、奈米乳液包埋藏紅花酸和固定化…………38
三、粒徑測量和包埋率測試實驗…………40
四、動物實驗…………45
伍.結論……………48
陸.參考文獻…………49

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