臺灣博碩士論文加值系統

中 文 摘 要
本論文研究目的，在探討具酪胺酸酶抑制活性之植物萃取物於美白化粧品之應用，以中草藥植物：桑 ( Mulberry )、甘草 ( Glycyrrhiza )、
牡丹皮 ( Tree Peony Bark ) 為材料，使用80 % 酒精、水為溶劑，進行微波萃取、減壓濃縮、冷凍乾燥，評估內容有: (一) 探討黑色素生成抑制活性﹔(二) 抗氧化生物活性﹔(三) 細胞存活率試驗評估﹔(四) 探討不同助溶劑之溶解度﹔(五)運用於保養產品應用調製。
在美白生物活性作用方面，結果顯示: ME (桑枝萃取液)、GE (甘草萃取液)、TE (牡丹皮萃取液)皆可對蘑菇酪胺酸酶的活性產生不同程度的抑制，其中以ME萃出物的抑制效果最佳﹔在B-16黑色素瘤細胞上清液之黑色素抑制試驗中ME萃出物的效果亦是較佳。將ME、GE、TE萃取液加入美白化粧水、精華液、乳液保養品配方中，其中以GE配方的蘑菇酪胺酸酶活性抑制較佳﹔在配方中加入4W，結果顯示以GE配方的蘑菇酪胺酸酶活性抑制較佳。
在抗氧化生物活性作用方面，運用清除DPPH自由基能力，結果顯示其中以TE萃出物抑制率較佳﹔運用清除ABTS陽離子自由基能力，結果顯示其中以TE萃出物抑制率較佳﹔其清除DPPH及ABTS自由基皆會隨著萃出物濃度增加而提高抑制率。
在細胞存活率方面，結果顯示: ME、GE、TE萃出物0.1mg/ml，皆有84 % 以上的細胞存活率﹔在0.01 mg/ml，皆有96 %以上的細胞存活率。
運用不同的助溶劑溶解，當助溶劑為Isoceteth-20其溶解力最佳，若與1-3 Bytlene glycol、Propylene glycol搭配使用，則溶解力增加，搭配使用量亦可相對減少。
最後，將ME萃取液加入化粧品配方中，並加入衛生署藥政處規範之部分美白成分調製，經臨床試驗，本論文所討論之精華液配方，對於某些自願者有良好的反應，經自願者連續使用六星期，其美白幼艦i由Lab值顯示，L數值增加達到1.5以上者，結果顯示實驗數據有良好的反應，可提供為良好化粧品原料之商業應用。

Abstract
The goal of the thesis is to evaluate the application of plant (Chinese medicine plant) extracts with potent tyrosinase inhibition activites on whitening cosmetics. The Mulberry, Glycyrrhiza, and Tree Peony Bark were employed as study materials. The selected plants were extracted by 80 % ethyl alcohols or distilled water by microwave machine, concentrated under a reduced pressure, and then dry by a lyophilizer. The evaluation of the application of herbal extracts on cosmetics includes: (1) suppression activities of melanin synthesis in vitro; (2) anti-oxidation activities; (3)cell survival rate; (4)solubility in cosmetic formula (5)efficacy in skin care products.
In the application on skin whitening, all these plants extracts, ME ( mulberry extract ), GE (Glycyrrhiza extract ), TE (Tree Peony Bark extract ), have the ability to inhibit the mushroom tyrosinase activities. Among them, the ME extract has the best inhibition activities. Similar effects were observed in B-16 melanoma cells. However, when combined with the known whitening ingredients, the GE extract expressed the highest inhibition activities.
In the anti-oxidation activity, TE extract contained a potent activity on the elimination of the DPPH free radical. Similar effect was observed in the TEAC experiment. We further examined the cytotoxicity of the plants extracts. All the materials showed little or none cytotoxicity at the concentration of 0.01 mg/ml.
These extracts have a good solubility in an Isoceteth-20 solution. Although 1-3 butylene glycol and propylene glycol were commonly used solvents, some precipitates were observed in these solutions.
Finally, we added the extracts together with four known whitening ingredients into the skin care formula, and evaluated their effects in vivo. Significant whitening effect was observed in some volunteers by a chromameter.

目 錄
中文摘要 …………………………………………………………………..I
英文摘要 ………………………………………………………………...III
誌謝 ………………………………………………………………………V
目錄 ………………………………………………………………….......VI
表目錄 …………………………………………………………………...IX
圖目錄 …………………………………………………………………...XI
縮寫對照表 …………………………………………………………..…XII

第壹章 緒論
1.1 前言 ………………………………………………….……1
1.2 文獻回顧 …………………………………………….……2
1.3 研究動機 …………………………………………….……6
第貳章 材料與方法
2.1 實驗架構 ………………………………………….………7
2.2 實驗藥材 …………………………………………………12
2.3 藥材成份簡介 ……………………………………………13
2.4 實驗藥品 …………………………………………………15
2.5 儀器設備 …………………………………………………17
2.6 實驗方法
2.6.1 樣品的製備 ………………………………………...……19
2.6.2 中草藥萃出物重量與原始濃度 ……………………..….20
2.7 中草藥萃取液之美白活性探討
2.7.1 中草藥萃出物對蘑菇酪胺酸酶活性抑制的測定 …...…21
2.7.2 中草藥萃出物對B-16黑色素瘤細胞上清液中黑色素的
抑制 ……………………………………………………..23
2.7.3 以電子顯微鏡觀察，中草藥萃出物對B-16黑色素瘤細胞
的影響 …………………………………………………..25
2.8 中草藥萃取液之抗氧化活性探討
2.8.1 Scavenging of 2,2-diphenyl-1-picryl-hydrazyl ( DPPH‧)
Assay …………………………………………………….26
2.8.2 Scavenging of Radical Cation 2,2-azinobis-( 3-ethylben-
zothiazoline-6-sulphonate ) ( ABTS‧+ ) assay …………27
2.9 中草藥萃出物之細胞存活率試驗
2.9.1 MTT assay ………………………………………………..29
2.10 中草藥萃取液不同助溶劑之溶解度探討
2.10.1 Mulberry Extract 不同助溶劑之溶解度 ……………….31
2.10.2 Glycyrrhiza Extract 不同助溶劑之溶解度 …………….32
2.10.3 Tree Peony Bark Extract 不同助溶劑之溶解度 ……….33
2.11 中草藥萃取液保養品配方中實際應用探討
2.11.1 中草藥萃取液保養品配方與劑型 ……………….……..34
2.11.2 中草藥萃取液保養品之人體試驗 …………….………..36
2.12 統計分析 ……………………………….………………...37
第叁章 結果
3.1 中草藥萃出物之重量 …………………………..………..38
3.1.1 中草藥萃取液之pH值 ……………………….…………39
3.2 中草藥萃取液美白活性試驗結果
3.2.1 中草藥萃出物對蘑菇酪胺酸酶活性的抑制率 …………40
3.2.2 中草藥萃出物對B-16黑色素瘤細胞上清液中黑色素
的抑制率 ………………………………………………...44
3.3 中草藥萃取液之氧化活性試驗結果
3.3.1 中草藥萃出物DPPH試驗結果 …………………………48
3.3.2 中草藥萃出物ABTS試驗結果 …………………………50
3.4 中草藥萃出物細胞存活結果 ……………………………51
3.5 中草藥萃取液不同助溶劑之溶解度試驗結果
3.5.1 ME不同助溶劑之溶解度試驗結果 …………………….52
3.5.2 GE不同助溶劑之溶解度試驗結果 …………………….53
3.5.3 TE不同助溶劑之溶解度試驗結果 …………………….55
3.6 中草藥萃取液在保養品配方中實際應用結果
3.6.1 ME萃取液保養品配方中人體試驗結果 ………………56
第肆章 討論 …………………………..………………………….65
第伍章 結論 …………………………………………..………….69
參考文獻 ………………………………………………………………...72







表 目 錄
表1、保養品配方成份含量 …………………………………………….33
表2、化粧水配方成分表 ……………………………………………….34
表3、精華液配方成分表 ……………………………………………….34
表4、乳液配方成分表 ………………………………………………….34
表5、以△E值表示色差分類等級 …………………………………….36
表6、ME、GE、TE 所得萃出物之重量 ………………………………37
表7、ME、GE、TE 萃取液之pH測定 ………………………………38
表8、ME、GE、TE萃出物對蘑菇酪胺酸酶活性的抑制 ……………39
表9、美白化粧水對蘑菇酪胺酸酶活性的抑制 ……………………….40
表10、美白精華液對蘑菇酪胺酸酶活性的抑制 ………………………41
表11、美白乳液對蘑菇酪胺酸酶活性的抑制 …………………………42
表12、ME、GE、TE萃出物對B-16黑色素瘤細胞上清液中黑色素的
抑制率 …………………………………………………………...43
表13、清除DPPH自由基之結果 ……………………………………..48
表14、清除ABTS自由基之結果 ……………………………………..49
表15、ME、GE、TE細胞存活率結果 ……………………………….50
表16、ME萃取液不同助溶劑之溶解度 ………………………………51
表17、GE萃取液不同助溶劑之溶解度 ………………………………53
表18、TE萃取液不同助溶劑之溶解度 ……………………………….54
表19、ML-A配方之人體試驗結果 ……………………………………55
表20、ML-B配方之人體試驗結果 ……………………………………56
表21、ML-C配方之人體試驗結果 ……………………………………56
表22、ML-D配方之人體試驗結果 …………………………………..57
表23、ME-A配方之人體試驗結果 …………………………………..58
表24、ME-B配方之人體試驗結果 …………………………………..59
表25、ME-C配方之人體試驗結果 …………………………………..59
表26、ME-D配方之人體試驗結果 …………………………………..60
表27、MM-A配方之人體試驗結果 ………………………………….61
表28、MM-B配方之人體試驗結果 ………………………………….62
表29、MM-C配方之人體試驗結果 ………………………………….62
表30、MM-D配方之人體試驗結果 ………………………………….63











圖 目 錄
圖1、實驗架構 ……………………………………………………………6
圖2、中草藥萃取液之美白活性探討 ……………………………...…....7
圖3、中草藥萃取液之抗氧化活性探討 …………………………………8
圖4、中草藥萃取液之細胞存活率試驗 …………………………….......9
圖5、中草藥萃取液不同助溶劑之溶解度試驗 ……………………….10
圖6、桑枝片、甘草、牡丹皮圖片介紹 ………………………………..11
圖7、ME不同濃度之B-16黑色素瘤細胞上清液中黑色素的抑制結果
………………………………………………………………….…44
圖8、GE不同濃度之B-16黑色素瘤細胞上清液中黑色素的抑制結果
…………………………………………………………………….45
圖9、TE不同濃度之B-16黑色素瘤細胞上清液中黑色素的抑制結果
…………………………………………………………………….46
圖10、美白化粧水配方產品使用六週之結果 ………………………. 57
圖11、美白精華液配方產品使用六週之結果 ………………………..60
圖12、美白乳液配方產品使用六週之結果 …………………………..63

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