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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/15179

Title: 90 % 甲醇或超臨界二氧化碳萃取雷公草活性成分之抗老化能力及其在化妝品的應用
The anti-aging capability of the active components extract from Centella Asiatica by 90 % methanol or supercritical CO2 and their application in cosmetic
Authors: Wen-Yeh Chen
陳玟燁
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: 雷公草;超臨界二氧化碳;人類皮膚纖維母細胞;膠原蛋白分泌
Centella asiatica;supercritical carbon dioxide;human skin fibroblast;collagen secretion
Date: 2008
Issue Date: 2011-06-30T08:07:58Z
Abstract: 本研究目的在探討用超臨界二氧化碳及傳統溶劑萃取法萃取之雷公草萃取物的抗老化能力與在抗皮膚老化化妝品上的應用。實驗分為兩部份:1、分別利用超臨界二氧化碳及甲醇萃取雷公草,進行萃取物之抗老化評估。在超臨界二氧化碳萃取方面先探討不同萃取溫度、壓力及助溶劑添加與否對萃取效果之影響,以求出最佳萃取條件,再與甲醇萃取物進行抗氧化能力比較,並以細胞培養方式觀察不同萃取方式所得之雷公草萃取物對人類纖維母細胞 ( CCD-966SK ) 存活率與促進膠原蛋白合成的影響。2、將上述實驗所得到之最佳萃取條件下的雷公草萃取物添加至乳霜基本配方中,並進行其特性之探討,包含安定性、安全性及有效性如皮膚保水性、彈性、粗糙度之評估。 實驗採用連續流動式之二氧化碳萃取雷公草中的有效成分,在溫度32 ~ 40 ℃,壓力15 ~ 45 MPa範圍下進行萃取,並比較添加助溶劑 對產率之影響,各萃取條件下之產物經由高效液相層析 ( HPLC ) 分析後,得到最佳萃取條件為35 ℃、35MPa、二氧化碳流速為 1 ml/min,並添加乙醇為助溶劑,其流速為0.1 ml/min。此條件下所得之萃取物中皆含有asiaticoside、madecasside、asiatic acid 及madecassic acid四種有效成份,與 90 % 甲醇萃取物層析圖譜比較後,顯示使用甲醇萃取可得到較多有效性成分。 以添加助溶劑之超臨界二氧化碳 (SFE-CO2) 萃取物與甲醇萃取物進行抗老化評估。在抗氧化試驗方面,兩種萃取物在濃度10 mg/mL下皆有良好的清除DPPH自由基能力,清除率可達90 % 以上;還原力以SFE-CO2萃取物效果較甲醇萃取物好,濃度為5 mg/mL時其還原力吸光值即與trolox無顯著差異;螯合亞鐵離子方面,甲醇萃取物螯合Fe2+ 能力隨濃度的增加而增高,在濃度為10 mg/mL下約有75 % 之螯合能力,而添加助溶劑之SFE-CO2萃取物螯合能力較差,皆低於30 %。細胞試驗方面,兩種萃取物在濃度10 ~ 125 μg/mL 之範圍內可使CCD-966SK存活率上升,對其膠原蛋白的合成亦有促進作用。 含或不含雷公草萃取物之乳霜經離心30分鐘後不會發生相分離,在高溫、室溫、低溫恆溫和循環變溫中貯存兩個月以上皆可維持正常不變質,顯示出該產品具有良好的安定性。 塗抹含0.5 % 雷公草萃取物之乳霜後不會發生皮膚紅腫現象,也不影響皮膚pH值,且有些許保濕效果。使用五週後對皮膚之彈性值R2有增加之情形,而在皮膚粗糙度試驗方面,其皮膚鱗片度、平滑度與皺紋度指標值也有明顯降低趨勢。
The objectives of the study are to explore the antioxidation properties of the Centella asiatica extracts prepared by supercritical carbon dioxide extraction ( SFE-CO2 ) or by conventional methanol extraction and apply the extracts as anti-aging active ingredient in skin care cosmetics. The study includes two parts. The first part is to evaluate the anti-aging properties of Centella asiatica extracts prepared by the methods mentioned. In regards to SFE-CO2 extraction, different temperatures between 32 ~ 40 ℃, pressures between 15~45 MPa and co-solvent are parameters studied and to establish the best extraction conditions. The antioxidation properties, effect on the cell viability, proliferation and enhancing collagen secretion of human skin fibroblast (CCD-966SK) exerted by the different extraction methods. The second part is to prepare and characterize the creams containing Centella asiatica extracts. The characteristics studied includes storage stability, safety and the efficacy of creams. Water holding capacity, skin elasticity and skin roughness were parameters to evaluate the efficacy. The extracts obtained from different extract conditions were analyzed with HPLC to establish the best extract conditions and was at 35℃, 35 MPa, CO2 flow rate 1.0 mL/min and co-solvent (ethanol) flow rate 0.1 mL/min. The extracts containing active ingredient of asiaticoside, madecasside, asiatic acid and madecassic acid. 90 % methanol extract contained more active ingredients than that obtained from SFE-CO2 extraction. To evaluate the anti-aging of extracts by SFE-CO2 with co-solvent or 90% methanol, both extracts showed 90 % scavenging effect at 10 mg/mL. The reducing power of the extracts by SFE-CO2 with co-solvent was better than that by 90 % methanol. It showed no significant different with trolox at 5 mg/mL. The chelating ferrous ions capacity increased with increasing the concentration of 90 % methanol extract, is about 75 % at 10 mg/mL. The extracts by SFE-CO2 with co-solvent showed little chelating effect, is below 30 %. Cell line test showed that both extracts promoted the cell viability on CCD-966SK and enhancing the collagen secretion capability in the concentration range of 10~125 μg/mL. The creams containing either with or without Centella asiatica extracts were stable for centrifuging (3000 rpm, 30 min), as well as stable more than two months at high temperature (50C), room temperature (25℃), low temperature (4℃) and temperature swing (4℃-25℃-50℃) storage. These results showed the product had good stability. The safety test resulted in no erythema based on the Daize score test and did not change skin pH value and had a little water holding capacity in terms of electrical capacitance. Skin elasticity (R2) for those whom have applied the creams containing Centella asiatica extracts increased over time during five weeks test period. Skin roughness also decreased over time during five weeks test period and did show a significant difference for skin scaleness, skin smoothness and skin wrinkles index.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0M95320025
http://ntour.ntou.edu.tw/ir/handle/987654321/15179
Appears in Collections:[食品科學系] 博碩士論文

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