| 摘要: | 近年來，化粧品市場崇尚天然，以中藥萃取液作為化粧品功效性原料勢必成為一股趨勢，因此中藥萃取液應用在化粧品中，被期許具備一定的皮膚功效。當歸（Angelica sinensis）是已知具有優異的抗氧化的能力，但其對皮膚美白的效能卻眾說紛紜，因此本研究希望深入瞭解當歸萃取液應用在皮膚美白的確切價值。
本研究設計六大目標進行探討：第一目標 – 瞭解不同萃取方法之萃取率：以兩種不同溶劑（95%乙醇和純水）與四種不同的萃取方法（加熱迴流萃取法、微波萃取法、冷浸萃取法及超聲波震盪萃取法）分別進行萃取，並比較其萃取率；第二目標 – 活性評估分析：主要針對美白及抗氧化等功效進行評估，並從中判斷活性與萃取方式之相關性；第三目標 – 安全性評估：將具有最佳功效之當歸萃取液，利用皮膚刺激性試驗與細胞毒性試驗評估萃取液之安全性；第四目標 – 安定性評估：將具有最佳功效之當歸萃取液，進行長時間安定性、熱安定性及不同 pH 值安定性評估，以瞭解萃取液成為保養品原料之可行性；第五目標 – 人體美白實驗：將當歸萃取液添加於基礎面霜配方中，進行人體美白功效評估，以瞭解皮膚美白效能；第六目標 – 活性成分之追蹤：將當歸萃取物進一步分離及純化，追蹤分析其具有美白效能之活性成分。
研究結果顯示，第一目標 - 瞭解萃取效率：水為溶劑所得之萃取物產率皆優於95%乙醇；第二目標 - 活性評估分析：95%乙醇對於酪胺酸酶的活性及黑色素生成上皆有抑制的能力，效果優於水萃物。而在抗氧化方面，95%乙醇在清除 DPPH． 和 ABTS．＋ 自由基能力也皆優於水萃物，其中以微波萃取方法效果為佳，在400 μg/mL時，清除率皆達80%；第三目標 - 安全性評估：細胞毒性經過 24 和 72 小時測試後，細胞存活率皆在80% 以上，在皮膚刺激性實驗中，結果也顯示當歸萃取液無皮膚刺激現象；第四目標 - 安定性評估：在長時間安定性、熱安定性和不同 pH 值的測試結果顯示，萃取液仍維持安定狀態。第五目標 - 人體美白實驗：經人體膚質試驗後，顯示具有優異的美白功效，能提高其 ITA° 值。第六部分 - 活性成分之追蹤：經由分離純化出的化合物 1，藉由1H-NMR、13C-NMR、薄層色層分析（TLC）圖譜解析以及相關文獻資料加以比對，確定為阿魏酸。在活性測試結果顯示，化合物 1 具有抗氧化能力及皮膚美白功效，故進一步證實當歸萃取添加於化粧品中，能有效減少黑色素之產生。
In the recent years, the cosmetics markets tends toward nature, therefore, using the extracts of natural plants to make as the ingredients of cosmetics has become a tendency, but the extract applications in cosmetics was hoped to have certain skin efficacy. Angelica sinensis is known as the excellent antioxidant, but opinions are widely divided on skin whitening. Therefore, in this study, we hope to understand the price of the applications of the A. sinensis extract in skin care.
In this study, we were divided into six major objectives to explore. The first objective, to understand the extraction yield of extractions in different extractive methods : used two different solvents ( 95% ethanol and H2O ) and four different extraction methods (such as refluxing, microwave, immersed, ultrasound) to calculate and compare yields of extractions, respectively. Secondly, analysis of activity assessment : the evulation of efficacy was mainly aimed at whitening and antioxidation, then, to determine the correlation between activity and extractions ; Thirdly, evaluation of safety : we selected the best efficacy extract, then used skin irritation and cytotoxicity test to assess the security of the extract ; Fourthly, the valuation of stability : we selected the best efficacy extract, and then used long time stability, thermal stability and different pH stability to assess stability of the extract become the skin care ingredient ; The fifth, human whitening experiment : in order to understand the skin whitening efficacy, we added the A. sinensis extract to the base cream, and it was evaluated by human whitening efficacy test. Finally, to track the active ingredient : we separated and purified the A. sinensis extract further, and then traced the active ingredient that have whitening efficacy.
The results show that the first objective - to understand the extraction efficiency: the extraction yield of water both better than 95% ethanol; Secondly, analysis of activity assessment : 95% ethanol extract both had significant inhibition on the tyrosinase activity and melanin production, which was better than the water extract. On the antioxidant activity, the 95% ethanol extraction had significant scavenging on the DPPH. and ABTS. + free radical capacity, the effect also better than the water extraction. Among, the microwave extraction had the best effect which the scavenging were over 80% at a concentration of 400 μg/mL. Thirdly - Evaluation of safety :On the cytotoxicity test at 24 and 72 hrs, the cell survival were more than 80%. In addition, the skin irritation test also showed that the extracts had no stimulation. Fourthly - Evaluation of stability : at long time stability, thermal stability and different pH test results both show that extracts still remained stable state. Fifth - Human whitening experiment : after human skin test, the result showed that the extract had excellent whitening effect, which can increase the ITA° values. Finally - Track the active ingredient : compounds 1 was through the separation and purification, which identified as ferulic acid by compare 1H-NMR, 13C-NMR, thin layer chromatography (TLC), analysis of chromatogram and related documents. On the activity test, compounds 1 had antioxidant capacity and skin whitening effect, therefore, we futhur confirmed that A. sinensis extract add to cosmetics, which can reduce the production of melanin effectively . |
