臺灣博碩士論文加值系統

本研究的目的在探討緬甸常見藥用植物黃香楝(Hesperethusa crenulata (Roxb.) Roem)在抗老化與美白化妝品上的有效性。在抗老化化學性試驗方面，於濃度範圍1－10 mg/mL不論是清除DPPH、超氧陰離子及過氧化氫上，黃香楝心材萃取物的抗氧化能力比樹皮萃取物佳。在美白化學性試驗方面，則以樹皮萃取物的酪胺酸?活性抑制效果較心材萃取物佳。在細胞株試驗方面，黃香楝心材萃取物及樹皮萃取物於1－1000 μg/mL時，都有促進纖維母細胞 CCD-966SK 增生及膠原蛋白生成的效果。而對於黑色素腫瘤細胞 A375的影響，樹皮萃取物於濃度超過 125 μg/mL以上時，其存活率開始有下降趨勢，而心材萃取物即使濃度達1000 μg/mL，對其存活率仍無影響。此外，樹皮及心材萃取物皆具有抑制黑色素腫瘤細胞生成黑色素的效果，以樹皮萃取物濃度為500 μg/mL時，對於UV-A照射後之黑色素腫瘤細胞生成黑色素可達到90 %的抑制效果。以黃香楝萃取物濃度1 %之乳霜試製品具有良好之安定性及皮膚安全性，而人體有效性試驗中，10位受試者塗抹含黃香楝萃取物乳霜5週後，其皮膚黑色素、鱗片度及皺紋度指標值有明顯降低趨勢，且均未出現皮膚刺激性或過敏現象。上述實驗結果證實，黃香楝之心材及樹皮萃取物不但具有顯著的抗老化與美白功效，未來更具有開發為功能性化妝品上之潛力。

Hesperethusa crenulata (Roxb.) Roem (family Rutaceae), a plant of traditional cosmetics in myanmar, is claimed to exert anti-aging and whitening efficacy. The antioxidant activity and whiting efficacy of H. crenulata stem bark extract and heart wood extract were evaluated by chemical tests, mushroom tyrosinase inhibition test and cultured melanocytes. H. crenulata heart wood extract (from 1 to 10 mg/mL) was superior to stem bark extract in scavenging effect on hydrogen peroxide , superoxide, DPPH free radicals. However, the inhibitory effect on mushroom tyrosinase showed that H. crenulata stem bark extract was superior to heart wood extract. Both extracts (from 1 to 1,000 μg/mL) increased CCD-966SK fibroblasts proliferation and collagen generation. In MTT test H. crenulata stem bark extract (above 125 μg/mL) decreased the viability of A375 human melanocytes, while heart wood extract (1,000 μg/mL) had no cytotoxic effect. Additionally, the inhibition effect by protecting from UV-A radiation was observed H. crenulata stem bark extract (500 μg/mL) decreased melanin formation by A375 cell by 90 %. Furthermore, the cosmetics cream products containing 1% H. crenulata stem bark or heart wood extract were with high stability and safety. Ten subjects applying the cream products significantly decreased in skin melanin, scaliness index, wrinkles index and melanin formation during five weeks test period, and none of them was observed skin irritability or allergy during test period. In conclusion, these results indicate that H. crenulata stem bark extract and heart wood extract have good anti-aging and whitening efficacy, and could be a potential ingredient to be applied in functional cosmetics in the future.

中文摘要………………………………………………….. Ⅰ
英文摘要………………………………………………….. Ⅱ
壹、 前言……………………………………………………….. 1
一、 黃香楝之藥用植物學考察………………………………….. 1
二、 黃香楝之植物學分類……………………………………….. 1
三、 黃香楝之已知成分………………………………………….. 1
貳、 文獻整理………………………………………………….. 2
一、 皮膚老化之簡介………………………………………….. 2
1. 皮膚老化的成因與變化………………………………….. 2
2. 影響皮膚老化的相關機制……………………………….. 3
3. 紫外線與色素沈澱之關係……………………………….. 5
4. 皮膚老化的自由基學說………………………………….. 6
二、 皮膚黑色素生成之簡介………………………………….. 6
1. 黑色素形成路徑………………………………………….. 7
2. 酪胺酸?………………………………………………….. 7
3. 美白劑之作用機制……………………………………….. 8
參、 材料與方法……………………………………………….. 9
一、 實驗大綱………………………………………………….. 9
二、 實驗流程………………………………………………….. 9
1. 黃香楝萃取物之抗老化與美白能力化學性試驗………. 10
2. 黃香楝萃取物之抗老化與美白能力細胞試驗…………. 10
3. 黃香楝萃取物乳霜的製作與其安定性、安全性測試及人體有效性試驗…………………………………………..
11
三、 實驗材料………………………………………………….. 12
四、 實驗方法………………………………………………… 16
1. 黃香楝萃取物之製備…………………………………….. 16
2. 基本成分分析…………………………………………….. 16
3. 細胞培養………………………………………………….. 19
4. 黃香楝萃取物抗老化能力評估………………………….. 20
4.1. 抗氧化成分之定量分析………………………………….. 20
4.1.1. 總酚類化合物含量(Total polyphenol)…………………... 20
4.1.2. 總類黃酮含量(Total flavonoid)………………………….. 21
4.2. 抗氧化活性測定………………………………………….. 22
4.2.1. 清除 DPPH 自由基能力之測定………………………... 22
4.2.2. 清除超氧陰離子能力之測定…………………………….. 23
4.2.3. 還原能力之測定………………………………………….. 24
4.2.4. 清除過氧化氫能力之測定.………………………………. 26
4.2.5. 黃香楝萃取物對促進纖維母細胞增生之作用…………. 27
4.2.6. 黃香楝萃取物對促進膠原蛋白合成之作用……………. 27
5. 黃香楝萃取物美白有效性試驗………………………….. 28
5.1. 黃香楝萃取液對於酪胺酸?活性抑制的影響…………. 28
5.2. 黃香楝萃取物對黑色素腫瘤細胞存活率之影響……… 29
5.3. 黃香楝萃取物對抑制黑色素腫瘤細胞生成黑色素影響 29
6. 含黃香楝萃取物之乳霜之製備………………………….. 29
6.1. 品質測試………………………………………………….. 30
6.2. 安定性測試………………………………………………. 30
6.3. 安全性測試……………………………………………….. 31
6.4. 有效性測試……………………………………………...... 31
7. 統計分析………………………………………………….. 33
肆、 結果與討論……………………………………………….. 34
一、 黃香楝萃取物之特性…………………………………….. 34
1. 萃取量與萃取率………………………………………….. 34
2. 基本成分方析…………………………………………….. 34
二、 黃香楝萃取物抗老化能力評估………………………….. 34
1. 抗氧化成分之定量分析………………………………….. 34
1.1. 總酚類化合物含量……………………………………….. 34
1.2. 總類黃酮含量…………………………………………….. 34
2. 抗氧化活性測定………………………………………….. 35
2.1. DPPH自由基之清除能力………………………………... 35
2.2. 超氧陰離子之清除能力………………………………….. 35
2.3. 還原力能力……………………………………………….. 36
2.4. 過氧化氫之清除能力…………………………………….. 36
3. 黃香楝萃取物之抗氧化細胞實驗………………………. 36
3.1. 黃香楝萃取物對促進纖維母細胞存活率之影響………. 37
3.2. 黃香楝萃取物對促進膠原蛋白合成之作用……………. 37
三、 黃香楝萃取物美白有效性試驗………………………….. 37
1. 黃香楝萃取物美白有效性試驗體外實驗………………. 37
1.1. 酪胺酸?活性之抑制…………………………………….. 37
2. 黃香楝萃取物美白有效性試驗細胞實驗………………. 38
2.1. 黃香楝萃取物對黑色素腫瘤細胞存活率之影響………. 38
2.2. 黃香楝萃取物對抑制黑色素腫瘤細胞生成黑色素影響 38
四、 含黃香楝萃取物之乳霜品質與人體試驗分析…………. 39
1. 品質特性………………………………………………….. 39
1.1. 安定性…………………………………………………….. 40
1.2. 色度測試………………………………………………….. 40
2. 對皮膚之安全性………………………………………….. 40
2.1. 皮膚敏感性……………………………………………….. 40
2.2. 皮膚 pH 值之測定………………………………………. 41
3. 有效性測試……………………………………………….. 41
3.1. 皮膚電容增加率（保水性）之 in vivo 測定…………… 41
3.1.1. 皮膚電容增加率短效性試驗…………………………….. 41
3.1.2. 皮膚電容增加率長效性試驗…………………………….. 41
3.2. 對皮膚美白效果之 in vivo 測定……………….............. 42
3.3. 對皮膚彈性效果之 in vivo 測定……………………….. 42
3.4. 對皮膚鱗片度效果之 in vivo 測定…………………….. 43
3.5. 對皮膚平滑度效果之 in vivo 測定…………………….. 43
3.6. 對皮膚皺紋度效果之 in vivo 測定…………….............. 43
伍、 結論……………………………………………………….. 45
陸、 參考文獻………………………………………………….. 46
表…………………………………………………………... 53
圖………………………………………………………….. 66
附錄……………………………………………………….. 91

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