Supercritical Carbon Dioxide Antisolvent Coating Propolis in Sucrose Palmitate Acid Ester to Increase Water Solubility

本研究以超臨界二氧化碳抗溶結晶(supercritical carbon dioxide antisolvent, SAS)共沉澱棕櫚酸蔗糖酯(sucrose palmitic acid esters, SE)及蜂膠，產製易溶水的蜂膠包覆物。首先探討超臨界抗溶壓力(120、140 bar)、溫度(33 oC)、抗溶時間(10、20 min)、進料流速(0.2、0.5、0.8 ml/min)及蜂膠、SE 之進料量比(3、4)對包覆物之總產率、單位載藥量及類黃酮回收率的影響。結果發現進料流速以 0.5 ml/min 可得較佳之總產率、載藥量及類黃酮回收率，抗溶壓力與時間對於包覆總產率、載藥量及類黃酮回收率影響不大，但進料量比確有很大差異。因此應用兩變數的應答曲面實驗設計(response surface methodology, RSM)尋找SAS 之溫度(33、43、53 oC)及進料濃度比(3、4、5)之最適化條件，而SAS 操作條件為壓力 120 bar、時間 10 min、進料流速 0.5 ml/min。結果 RSM 預測的最佳應答值為進料濃度比 3.71、溫度 53oC，可得包覆總產率 55.4%、載藥量 22.9% 及類黃酮回收率 54.4%，此與實驗值相近。由 RSM 結果發現，抗溶溫度影響不大，但進料比值，對 SAS 包覆物的總產率、載藥量及類黃酮回收率影響較大。pH 7.4 模擬腸液之溶離度實驗結果顯示，包覆後的蜂膠溶解效果比蜂膠丙酮萃取物佳，表示較溶水的棕櫚酸蔗糖酯與蜂膠共沉澱後產生連接，更能增加蜂膠之溶水性。應用 SAS 共沉澱棕櫚酸蔗糖酯及蜂膠能產製易溶水之蜂膠，以提升蜂膠在保健食品之應用性。In this study the supercritical carbon dioxide antisolvent (SAS) co-precipitation method is applied to sucrose palmitic acid esters (SE) and propolis to produce water soluble propolis coating. The first step is to investigate the impacts of supercritical antisolvent pressure (120, 140 bar), temperature (33oC), time (10, 20 min), feed flow rate (0.2, 0.5, 0.8 ml/min) and the feed concentration ratio between propolis and SE (3, 4) on the total yield, drug content, and flavonoids recovery rate of the coating. The results revealed that the feed flow rate of 0.5 ml/min has led to better total yield, drug content, and flavonoids recovery rate. The antisolvent pressure and time have not led to significant impacts on total total yield, drug content, and flavonoids recovery rate, yet there is a significant difference in the feed concentration ratio. Therefore the bivariate response surface methodology (RSM) has been applied to obtain the optimal conditions of temperature (33, 43, 53 oC) and feed concentration ratio (3, 4, 5) of SAS with operation conditions of pressure at 120 bar, time at 10 min, and feed flow rate at 0.5 ml/min. The results indicate that the optimal responses predicted by RSM are the feed concentration ratio of 3.71 and the temperature of 53oC, which lead to the total yield of 55.4%, the drug content of 22.9%, and the flavonoids recovery rate of 54.4%, all of which are similar to the experimental values. The RSM results reveal that antisolvent temperature does not lead to any significant impact, while the feed concentration ratio can result in more significant impact on the total yield, drug content, and the flavonoids recovery rate of SAS coating. The result of dissolution rate experiment based on pH 7.4 simulated intestinal fluids indicated that coated propolis dissolved better than acetone extracts of propolis. This indicates that the bonding generated from the co-precipitation between water soluble SE and propolis can enhance the solubility of propolis. Thus the SAS co-precipitation of SE and propolis can be applied to produce water soluble propolis in order to improve the applicability of propolis among health foods.摘要 I Abstract III 縮寫表 V 目錄 VI 表目錄 VIII 圖目錄 IX 附表目錄 X 第一章 緒論 1 1-1 研究動機 1 1-2 研究目的與規劃 2 第二章 文獻回顧 4 2-1 蜂膠介紹 4 2-3 乳化劑介紹 6 2-4 蔗糖酯介紹 8 2-5 超臨界二氧化碳的技術與應用 9 2-6 反應曲面法介紹 14 第三章 實驗材料與方法 15 3-1 原料與材料製備 15 3-2 試劑與藥品 15 3-2-1 氣體:二氧化碳 15 3-2-2 藥品 16 3-3 實驗設備 18 3-3-1 超音波攪拌萃取設備 18 3-3-2 索式萃取設備 19 3-3-3 超臨界抗溶結晶設備 20 3-3-4 紫外-可見分光光度儀 24 3-3-5 示差掃描熱分析儀 24 3-3-6 場發射掃描式電子顯微鏡 24 3-3-7 粒徑分析儀 25 3-3-8 其他設備 25 3-4 實驗方法與步驟 27 3-4-1 超音波攪拌萃取蜂膠之類黃酮 27 3-4-2 索式萃取蜂膠之類黃酮 27 3-4-3 棕櫚酸蔗糖酯與蜂膠萃取物在溶劑的溶解度實驗 28 3-4-4 總類黃酮含量測定及檢量線製作 28 3-5 超臨界抗溶沉澱過程 29 3-6 溶離度實驗 32 3-7 粒徑分析 33 3-8 電子顯微鏡(SEM)分析 33 第四章 結果與討論 34 4-1 蜂膠類黃酮萃取實驗 34 4-2 蜂膠與棕櫚酸蔗糖酯在溶劑的溶解度實驗 37 4-3 超臨界二氧化碳抗溶包覆蜂膠的預實驗 39 4-3-1 進料流速(QF)的影響 39 4-3-2 時間(t)的影響 39 4-3-3 壓力(P)的影響 40 4-3-4 進料量比(Wpro/WSE)的影響 40 4-4 應答曲面設計 SAS 包覆蜂膠實驗 42 4-4-1 包覆總產率(TY)應答分析值 45 4-4-2 載藥量(DC)應答分析值 45 4-4-3 類黃酮回收率(R)應答分析值 46 4-4-4 RSM的預測值與實驗值比較 46 4-5 溶離度 48 4-6 示差掃描熱分析儀(DSC) 50 4-7 粒徑分析 51 4-8 電子顯微鏡(SEM)共沉澱物晶貌 52 第五章 結論 55 參考文獻 6