臺灣博碩士論文加值系統

本研究目的探討，利用不同酸從烏骨雞雞爪與雞爪皮萃取含黑色素之膠原蛋白，並以醋酸所萃取白肉雞雞爪之膠原蛋白作為對照組，測定膠原蛋白濃度、產率與真黑色素含量的多寡，並測其UV光譜、抗氧化能力、吸濕率與抑制黑色素能力等特性，再以效果最佳者製成面霜，測定面霜於儲藏其間之物理與乳化安定性，並塗抹於8位20∼30歲自願者內側手臂，測量正常及誘發乾性皮膚角質層之水合量與正常皮膚水份散失量之成效。
研究結果顯示：經不同酸萃取烏骨雞爪及雞爪皮之膠原蛋白，由醋酸及鹽酸萃取者含有顯著較高之膠原蛋白濃度(p<0.05)且雞爪與雞爪皮間無顯著性差異，並發現醋酸與乳酸萃取之膠原蛋白有較佳之產率。真黑色素含量，則以醋酸萃取烏骨雞雞爪皮(SA)效果最佳；經不同酸萃取烏骨雞雞爪與雞爪皮之膠原蛋白，相較於白肉雞雞爪之膠原蛋白皆具有良好的吸收光譜。在不同相對濕度的吸濕率方面，相同相對濕度下，不同酸萃取之膠原蛋白，無顯著性的差異，但隨著相對濕度的提高其有顯著性的增加(p<0.05)。烏骨雞雞爪與雞爪皮膠原蛋白於1 mg/mL時，以醋酸萃取烏骨雞雞爪皮顯著具較佳的清除超氧陰離子與螯合亞鐵離子能力(p<0.05)且隨著濃度的提高其抗氧化力亦隨之增加。於抑制酪胺酸酉每與Dopachrome等抑制黑色素形成方面，亦以醋酸萃取烏骨雞雞爪皮顯著具較佳的效果(p<0.05) 且隨著濃度的提高其抑制率亦隨之上升。由效果最佳的醋酸萃取烏骨雞雞爪皮膠原蛋白製成面霜，並以基劑作為對照組，儲藏於25 ℃與50 ℃下，發現其物理安定性隨儲藏天數的增加，其表面黏性與彈性於0-14天皆有下降趨勢，但14天後漸趨平緩。在乳化安定性方面，儲藏於25 ℃或50 ℃下，經三個月以上皆無分離的現象。在面霜保濕性測定方面，經皮膚水份蒸散測定儀測定皮膚水份散失量的變化中發現，醋酸萃取烏骨雞雞爪皮膠原蛋白面霜與市售膠原蛋白面霜，相較於基劑皆顯著降低皮膚水分散失(p<0.05)且效果可達兩個小時以上。於正常皮膚角質層之水合量與誘發乾性皮膚角質層水合量方面，醋酸萃取烏骨雞雞爪皮膠原蛋白面霜其相較於基劑與市售膠原蛋白面霜皆有顯著性提高皮膚角質層水合量(p<0.05)效果可達兩小時以上。

The purpose of this study is to investigate the collagen content, production rate, and eumelanin content on collagen extracted from silky fowl feet and feet skin by various acids. The collagen extracted from broiler feet by acetic acid as the contractor and measured collagens’ UV spectrum, anti-oxidation ability, moisture absorption rate, and melanin inhibition capability. The cream samples were made by the extracted collagen has the best measurement results. Among all creams spreading on the inner side of arms of volunteers, whose are between age 20 to 30-year-old, and measure skin surface hydration on normal or bring out dry skin and skin surface transepidermal water loss on normal skin.
The results showed that the collagen extracted by acetic acid and hydrochloric acid have significantly higher collagen contents (p<0.05), but there is no significant difference between extract sources, feet and feet skin. However, extraction of both using acetic acid and lactic acid have higher collagen production rate. For eumelanin content test, collagen extracted from the skin of silky fowl feet by acetic acid (SA) have a better result and also find that collagen extracted from feet and feet skin of silky fowl by different acids have a better UV absorb results, comparing to collagen extracted from broiler feet. In moisture absorption rate test, collagens extracted by different acids have no significant difference on results in the same relative humidity. However, the difference is significantly (p<0.05) increasing with increasing of relative humidity. At 1 mg/mL concentration, collagen extracted from silky fowl feet skin by acetic acid has a significantly better ability of superoxide anion clearance and ferrous ion chelating ability (p<0.05) and the anti-oxidation ability increases with increasing of collagen concentration. Tests for Tyrosinase and Dopachrome inhibit ability (melanin producing inhibit ability), collagen extracted from skin of silky fowl feet by acetic acid also has a significant better result (p<0.05) and the inhibition rate increases with increasing of collagen concentrations and select collagen that has the best test result, using collagen extracted from the skin of silky fowl feet by acetic acid, to make creams and then store along with the contract examples, vehicle, at 25 ℃ and 50 ℃. The results showed the physical stability is increased during storage period but the apparent viscosity and elasticity is slightly decreased. For the emulsion stability test, all products will still keep in shape after three months or longer storage. For skin moisture retention test, using Tewameter TM210 to exam the skin moisture change, it found out that cream made by collagen extracted from silky fowl feet skin by acetic acid and the cream sold on the market have a significant effect on decreasing skin surface transepidermal water loss (p<0.05), comparing to vehicle, and the effect can persisted longer then 2 hours. Normal skin surface for hydration test, cream made by the collagen which is extracted from skin of silky fowl feet by acetic acid has a significantly better effect on increasing skin surface hydration (p<0.05), comparing to vehicle and commercial collagen cream, and the effect can persist longer than two hours. For the test of surface hydration on bring out dry skin has the same conclusion that cream made by collagen which is extracted from the skin of silky fowl feet by acetic acid has a significantly better effect on increasing skin surface hydration (p<0.05), comparing to vehicle and commercial collagen cream.

壹、中文摘要 I
貳、英文摘要 III
參、前言 1
肆、文獻探討 3
一、烏骨雞簡介 3
二、皮膚構造 3
(一)表皮層( epidermis ) 3
1.角質細胞( keratinocyte ) 4
2.黑色素細胞( melanocyte ) 4
3.蘭格罕氏細胞( langerhans cell ) 5
(二)真皮層( dermis ) 5
(三)皮下組織( subcutaneous tissue ) 5
三、黑色素的形成 7
(一)黑色素細胞的分布 7
(二)酪胺酸酉每的特性 9
(三)黑色素的生物合成 9
(四)紫外線對黑色素的作用 12
四、黑色素生成的抑制作用 14
(一)酪胺酸酉每活性的抑制 14
(二)氧化競爭酪胺酸酉每 14
(三)黑色素的還原作用 14
五、自由基與抗氧化機制 16
(一)自由基與活性氧群 16
1.超氧陰離子自由基( superoxide anion radical, O2-． ) 16
2.過氧化氫( hydrogen peroxide, H2O2 ) 17
3.羥基自由基( hydroxy radical, •OH ) 17
4.單重態氧( 或稱激氧 singlet oxygen, 1O2 ) 17
5.脂質過氧化物( lipid peroxide, LPO ) 17
(二)抗氧化劑的作用原理與種類 18
1.自由基終止劑( free radical terminator ) 18
2.還原劑( reducing agent ) 18
3.金屬離子螯合劑( metal chelator ) 19
4.單重氧的抑制劑( singlet oxygen inhibitor ) 19
六、皮膚保濕作用 20
(一)皮膚狀態的評估法 20
1.皮膚表面形態 20
2.角質層含水量 20
3.穿皮失水量( transepidermal water loss；TEWL ) 21
4.角質化程度不全 21
(二)保濕劑(humectants)介紹 21
1.親水性的保濕劑 22
2.疏水性的保濕劑 22
七、膠原蛋白簡介 23
(一)膠原蛋白的結構 23
(二)膠原蛋白的生合成 25
(三)膠原蛋白之純化與重組 27
(四)膠原蛋白之分類 28
(五)膠原蛋白的交聯 30
(六)膠原蛋白之降解 30
(七)膠原蛋白的應用 31
1.膠原蛋白在生醫材料的應用 31
2.膠原蛋白在醫藥上的應用 31
3.膠原蛋白在化妝品上的應用 32
伍、材料與方法 34
一、試驗流程 34
二、實驗材料 35
三、一般成分分析 35
1.水分 35
2.粗脂肪 35
3.灰分 35
4.粗蛋白質 36
四、烏骨雞雞爪膠原蛋白分析 37
(一)膠原蛋白萃取 37
(二)膠原蛋白之含量測定 37
(三)真黑色素含量 40
(四)膠原蛋白電泳分析 41
1.試藥配製 41
2.膠片配方 43
3.電泳操作 43
(五)UV光譜 44
(六)產率( % ) 44
(七)膠原蛋白在不同濕度下的吸濕率測定 45
五、烏骨雞雞爪膠原蛋白抗氧化特性之探討 46
(一)共軛雙烯法之測定 46
(二)還原力之測定 46
(三)清除DPPH自由基 46
(四)螯合亞鐵離子能力之測定 47
(五)清除超氧陰離子能力之測定 47
六、烏骨雞雞爪膠原蛋白抑制黑色素形成之測定 48
(一)酪胺酸酉每抑制率 48
(二)Dopachrome抑制率 48
七、烏骨雞雞爪膠原蛋白面霜 50
(一)面霜之製備 50
(二)物理安定性研究 50
1.pH值之測量 50
2.物性測定儀之測量 51
3.乳化安定性之測量 51
4.離心分離試驗 51
(三)面霜色澤之測定 52
八、膠原蛋白面霜對保濕能力之測定 52
1.自願受測者的條件 52
2.溫濕度條件 52
3.測定樣品 52
(a)市售膠原蛋白面霜 52
(b)烏骨雞雞爪皮膠原蛋白面霜 52
(c)基劑 52
4.測定項目 53
(a)膠原蛋白面霜對正常皮膚角質層水合量的測定 53
(b)膠原蛋白面霜對誘發乾性皮膚角質層水合量的測定 53
(c)膠原蛋白面霜對於正常皮膚水份散失量的測定 53
九、統計分析 54
陸、結果與討論 55
一、一般成分分析 55
二、烏骨雞雞爪膠原蛋白分析 55
(一)膠原蛋白外觀 55
(二)膠原蛋白含量測定 58
(三)真黑色素含量測定 60
(四)膠原蛋白產率 62
(五)膠原蛋白電泳分析 64
(六)膠原蛋白於不同溼度下之吸濕率測定 66
(七)膠原蛋白之UV光譜 68
三、烏骨雞雞爪膠原蛋白抗氧化特性之探討 70
(一)清除超氧陰離子測定 70
(二)清除DPPH自由基測定 72
(三)還原力測定 74
(四)螯合亞鐵離子測定 76
(五)共軛雙烯測定 78
四、烏骨雞雞爪膠原蛋白抑制黑色素形成之測定 80
(一)抑制酪胺酸酉每能力測定 80
(二)抑制Dopachrome能力測定 82
五、膠原蛋白面霜之測定 84
(一)膠原蛋白面霜之外觀 84
(二)面霜色澤之測定 84
(三)面霜之物理與乳化安定性 86
六、膠原蛋白面霜保濕性能之測定 89
(一)面霜對正常皮膚水份散失量之測定 89
(二)面霜對正常皮膚角質層水合量之測定 91
(三)面霜對誘發乾性皮膚角質層水合量之測定 93
柒、結論 95
捌、參考文獻 97

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