臺灣博碩士論文加值系統

本研究以製備出天然無毒性、高保濕性與美白功效之複合面膜為主軸，使用高分子材料聚乙烯吡咯烷酮(polyvinyl pyrrolidone，PVP)、明膠(Gelatin，GEL)與桑白皮(Morus alba，MA)結合於水針布織布(Spunlace Nonwoven)進行製備，期許可以於化妝品、保養品方面作應用。
本研究實驗製程上可分為第一階段：天然中草藥藥物萃取與美白評估，第二階段：PVP/GEL/MA奈米纖維薄膜製備，第三階段：薄膜與不織布結合製備三階段。首先將天然中草藥國槐、雞血藤、桑白皮、玉竹分別浸泡於純水與95%乙醇溶劑中進行中草藥萃取，藉由中草藥萃取率、細胞相容性、細胞存活率來評估四種中草藥之特性，並經酪胺酸酶抑制率測試選出純水萃取之桑白皮(Morus alba，MA)萃取液為最佳美白萃取液。再利用靜電紡絲技術將最佳調配濃度之PVP/GEL/MA溶液噴塗於水針不織布製成PVP/GEL/MA複合面膜。
結果顯示中草藥水萃取桑白皮在濃度0.5 mg/ml具有良好之酪胺酸酶抑制率，且此藥物濃度對於L929細胞相容性皆無任何排斥性，細胞存活率平均達到88%。靜電紡絲薄膜顯示，PVP/GEL綜合濃度提升至20 wt %，混合比例達到80/20時呈現光滑平順之纖維型態，隨著明膠與桑白皮藥物的添加使溶液的黏度、電導度之提升，紡絲參數在20 kV、15 cm、0.5 mg/hr時達到幾乎無珠粒與纖維纖細之型態，纖維直徑達到210 nm。PVP/GEL/MA薄膜與水針布作結合後由水接觸角觀測中發現角度位在29.2°，表明有良好之親水性能。水氣透過率中顯示約600-800 g/m2day，而人體皮膚濕的情況下為350 g/m2day，表示貼附於臉部時有良好之透氣效果。由酪胺酸酶抑制率中發現藥物釋放25分鐘，釋放率75%，抑制率為81.42±1.6%，美白功效些許高於市售面膜，且PVP/GEL/MA複合面膜皆不妨礙細胞生長，細胞存活率達88.79%，表明擁有良好生物相容性。證實本研究成功製備PVP/GEL/MA複合面膜並具備良好之美白功效與生物相容性能。

This study proposes composite masks with avirulence, humectancy, and whitening properties. Polymeric materials, including polyvinyl pyrrolidone (PVP), gelatin (GEL), and Morus alba (MA), are combined with spunlace nonwoven fabrics in order to form cosmetic care products. This study can be divided into three stages. In the 1st stage： natural Chinese herbal medicine are extracted and evaluated for whitening efficacy. In the 2nd stage, PVP/GEL/MA nanofiber membranes are prepared. In the last stage, membranes are then combined with nonwoven fabrics. Natural Chinese herbal medicine, including Sophora japonica, Spatholobus suberectus, Morus alba, and Polygonatum odoratum are respectively immersed in pure water and 95% ethanol solution in order to have the Chinese herbal medicine extracts. The extracts are tested in terms of extractability, biocompatibility, and MTT assay, thereby characterizing the properties of the four Chinese herbal medicine. Tyrosinase inhibition ratio is used to examine the water-extraction Morus alba (MA) extract to be the optimal whitening extract. Moreover, the PVP/GEL/MA solution is electrospun into spunlance nonwoven, forming PVP/GEL/MA composite masks.
The test results show that MA herbal extract at a concentration of 0.5 mg/ml has good inhibition ratio against tyrosinase, and the MTT assay result is 88 %, indicating a good biocompatibility with L929 fibroblasts. The electrospinning film has a smooth fiber formation when a 20 wt% PVA/GEL mixture is used. The addition of GEL and MA increases both the viscosity and electrical conductivity of the mixtures. The electrospun nanofibers are slim without bead-shaped and have a diameter of 210 nm with the settings of a voltage of being 20kV, a collect distance of being 15 cm, and an electrospinning rate being 0.5 mg/hr. Based on the contact angle meter restuls, the combination of PVP/GEL/MA film and spunlance nonwoven results in a water contact angle of 29.2°, suggesting a good hydrophility. The water vapor transmission rate of PVP/GEL/MA mask is 600-800 g/m2day, which is close to that of the human skin (350 g/m2day), which means the masks have good air-permeable property and suitable for the skin. Moreover, according to the tyrosinase inhibition measurement, the masks have a drug delivery period of 25 minutes at a release rate of 75 %, and an inhibition ratio of 81.42±1.6%. Namely, the whitening effect of the proposed composite masks is slightly higher than that of ready-made masks. In addition, the proposed composite masks have an MTT assay result at 88.79%, which shows that PPVP/GEL-MA composite masks does not obstruct the cell growth and have good biocompatibility. All test results confirm that the PVP/GEL/MA composite membranes have good whitening efficacy and biocompatibility.

中文摘要 I
ABSTRACT III
目錄 V
第一章 緒論 1
1.1前言 1
1.2研究背景 3
1.2.1國槐(Sophora japonica) 3
1.2.2雞血藤(Spatholobus suberectus) 4
1.2.3桑白皮(Morus alba) 5
1.2.4玉竹(Polygonatum odoratum) 6
1.3高分子材料 7
1.3.1聚乙烯吡咯烷酮(polyvinyl pyrrolidone) 7
1.3.2明膠(Gelatin) 8
1.4皮膚 9
1.4.1皮膚的構造 9
1.4.2皮膚的層次(Layers of the skin) 9
1.4.3皮膚的功能 14
1.5黑色素細胞(Melanocytes) 15
1.5.1黑色素的種類： 16
1.5.2黑色素(Melanin)的生成： 17
1.6酪胺酸酶(Tyrosinase) 19
1.6.1酪胺酸酶活性與人類皮膚之關係 20
1.6.2酪胺酸酶影響皮膚顏色深淺之決定因子 21
1.7紫外線(Ultraviolet light) 22
1.8美白成分與美白效果評估方法 24
1.8.1衛生福利部核定之美白成分 24
1.9靜電紡絲(electrospinning) 25
1.9.1靜電紡絲設備 26
1.9.2靜電紡絲過程之相關研究 27
1.10水針不織布(Spunlace Nonwoven) 28
1.11研究動機 29
1.12研究目的 31
1.13文獻回顧 33
1.14國內外相關專利文獻回顧 39
第二章 原理 52
2.1靜電紡絲原理 52
2.2掃描式電子顯微鏡原理 53
2.3減壓濃縮原理 54
2.4水接觸角原理 55
2.5專有名詞解釋 55
第三章 實驗 57
3.1實驗流程圖 57
3.1.1中草藥萃取液製備流程圖 57
3.1.2聚乙烯吡咯烷酮/明膠/桑白皮複合電紡薄膜製備流程圖 60
3.2實驗材料 65
3.3實驗設備 66
3.4實驗參數設計 67
3.4.1靜電紡絲加工參數設計 67
3.4.2中草藥國槐、雞血藤、桑白皮、玉竹實驗參數設計 67
3.4.3複合靜電紡絲薄膜實驗參數設計 67
3.5中草藥國槐、雞血藤、桑白皮、玉竹測試項目 68
3.5.1萃取率測試 68
3.5.2紫外分光光度計測試 68
3.5.3細胞相容性測試 68
3.5.4細胞存活率測試(MTT Assay) 69
3.5.5酪胺酸酶抑制性測試 71
3.6複合電紡薄膜測試項目 73
3.6.1複合電紡薄膜形態觀察 73
3.6.2纖維直徑(fibre diameter)分析測試 73
3.6.3傅立葉轉換紅外線光譜(FT-IR) 73
3.6.4藥物釋放測試 74
3.7複合面膜之測試項目 75
3.7.1水接觸角測試 75
3.7.2水分擴散測試 76
3.7.3水氣透過率測試 76
第四章 結果與討論 78
4.1國槐、雞血藤、桑白皮、玉竹萃取液其吸收度與萃取率評估 78
4.2中草藥國槐、雞血藤、桑白皮、玉竹萃取物對酪胺酸酶之抑制率 85
4.3中草藥國槐、雞血藤、桑白皮、玉竹萃取物對細胞相容性之影響 89
4.4中草藥國槐、雞血藤、桑白皮、玉竹萃取物對細胞存活性之影響 97
4.5 PVP、GEL與MA混合溶液之黏度與電導度 98
4.6不同濃度之PVP、PVP/GEL與PVP/GEL/MA複合電紡薄膜之掃 描式電子顯微鏡觀察 102
4.7 PVP/GEL/MA複合電紡薄膜之傅立葉轉換紅外線光譜 114
4.8 PVP/GEL/MA複合電紡薄膜之藥物釋放 115
4.9 PVP/GEL/MA複合電紡薄膜對細胞相容性之影響 118
4.10 PVP/GEL/MA複合電紡薄膜對細胞存活性之影響 124
4.11 PVP/GEL/MA/SN複合面膜之水接觸角 125
4.12 PVP/GEL/MA/SN複合面膜之水分擴散 127
4.13 PVP/GEL/MA/SN複合面膜之水氣透過率 129
4.14 PVP/GEL/MA/SN複合面膜之酪胺酸酶抑制率 130
4.15 PVP/GEL/MA/SN複合面膜之細胞存活率 132
4.16本論文研究成果在工業製品的應用 133
第五章 結論 134
第六章 建議 137
參考文獻 139

[1] 陳玉姬. 從資源基礎觀點探討化妝品市場之品牌, 通路與價格的競爭優勢—以台灣化妝品市場為例. 明道大學管理學院企業高階管理碩士班碩士論文.
[2] 鄭慈千. 化妝保養品市場新趨勢研究. 台北城市大學學報 2013；第36期：p.214-22.
[3] 吳普, 孫星衍, 張仲景, 孫馮翼, 梅彪. 神農本草經： 商務印書館； 1937.
[4] Lo Y-H, Lin R-D, Lin Y-P, Liu Y-L, Lee M-H. Active constituents from Sophora japonica exhibiting cellular tyrosinase inhibition in human epidermal melanocytes. Journal of ethnopharmacology 2009；124：625-9.
[5] 叶橘泉. 现代实用中药. 科技卫生出版社； 1958.
[6] Li W, Liu J, Guan R, Chen J, Yang D, Zhao Z, et al. Chemical characterization of procyanidins from Spatholobus suberectus and their antioxidative and anticancer activities. Journal of Functional Foods 2015；12：468-77.
[7] Yu Z-R, Hung C-C, Weng Y-M, Su C-L, Wang B-J. Physiochemical, antioxidant and whitening properties of extract from root cortices of mulberry as affected by membrane process. LWT-Food Science and Technology 2007；40：900-7.
[8] 李时珍. 本草綱目： 上海古籍出版社； 1991.
[9] Kang KB, Lee DY, Kim TB, Kim SH, Kim HJ, Kim J, et al. Prediction of tyrosinase inhibitory activities of Morus alba root bark extracts from HPLC fingerprints. Microchemical Journal 2013；110：731-8.
[10] 徐雅淳. 桑白皮萃取物之抗氧化活性及其對中波長紫外線輻射傷害之保護作用. 2008.
[11] 陈嘉谟, 王淑民, 陳湘萍, 周超凡. 本草蒙筌： 人民衛生出版社； 1988.
[12] Zhou X, Yuping Z, Zhao H, Liang J, Zhang Y, Shi S. Antioxidant homoisoflavonoids from Polygonatum odoratum. Food chemistry 2015；186：63-8.
[13] Liu X, Zhang M, Guo K, Jia A, Shi Y, Gao G, et al. Cellulase-assisted extraction, characterization, and bioactivity of polysaccharides from Polygonatum odoratum. International journal of biological macromolecules 2015；75：258-65.
[14] https：//zh.wikipedia.org/wiki/%E8%81%9A%E4%B9%99%E7%83%AF%E5%90%A1%E5%92%AF%E7%83%B7%E9%85%AE.
[15] Yan H. The hydrophobic zeolite molecular sieves and its application in CO_2 control technology. Shaanxi Chemical Industry 2002；3：005.
[16] 鄭棋文. 聚乙烯吡咯烷酮/聚醚胺混摻物之合成, 性質及應用研究 Part 2： 於乳化柴油製備之研究. 2012.
[17] 柯佩欣. 聚乙烯吡咯烷酮/聚醚胺混摻物之合成, 性質及應用研究 Part1： 合成與性質研究. 2012.
[18] 黃連佑. 明膠/CMC複凝聚製備尤加利精油微膠囊之研究. 正修科技大學 2014.
[19] 林育勤. 以明膠或明膠-酚醛樹脂混摻體為模板合成中孔洞材料. 2007.
[20] 許延年、蔡文玲、邱品齊、石博宇、周彥吉、黃宜純. 醫學美容 Medical Cosmetology. 華杏出版機構 2011.9：19.
[21] Pamela Hill RN. 美容師依定要會的-換膚與化學去角質. 合記圖書出版社 2012.1：36.
[22] http：//angelialin.pixnet.net/blog/post/285001325-%E3%80%90%E7%BE%8E%E5%AE%B9%E7%AD%86%E8%A8%98%E3%80%91%E7%9A%AE%E8%86%9A%E7%9A%84%E6%A7%8B%E9%80%A0%E3%80%81%E7%B4%8B%E7%90%86%E3%80%81%E9%99%84%E5%B1%AC%E5%99%A8%E5%AE%98.
[23] 洪勖峰. 槐花萃取物的美白研究. 中國醫藥大學中西醫結合研究所碩士論文 台中, 台灣 曹智莞 2003.
[24] 林志昇. 不同靈芝菌絲體萃取物對酪胺酸酶抑制活性之影響. 2010.
[25] http：//www.bjhuancai.com/news/html/?386.html.
[26] 朱志維. 利用麴酸抑制黑色素細胞生成與釋放黑色素研究進而建立一套美白機制. 國立暨南國際大學 2006.
[27] Prota G. Recent advances in the chemistry of melanogenesis in mammals. Journal of Investigative Dermatology 1980；75：122-7.
[28] Goldsmith LA. Biochemistry and Physiology of the Skin： Oxford University Press； 1991.
[29] Sánchez-Ferrer Á, Rodríguez-López JN, García-Cánovas F, García-Carmona F. Tyrosinase： a comprehensive review of its mechanism. Biochimica et Biophysica Acta (BBA)-Protein Structure and Molecular Enzymology 1995；1247：1-11.
[30] Mirica LM, Vance M, Rudd DJ, Hedman B, Hodgson KO, Solomon EI, et al. Tyrosinase reactivity in a model complex： an alternative hydroxylation mechanism. Science 2005；308：1890-2.
[31] Sugimoto K, Nishimura T, Nomura K, Sugimoto K, Kuriki T. Inhibitory Effects of. ALPHA.-Arbutin on Melanin Synthesis in Cultured Human Melanoma Cells and a Three-Dimensional Human Skin Model. Biological and Pharmaceutical Bulletin 2004；27：510-4.
[32] Enculescu M, Evanghelidis A, Enculescu I. Influence of morphology on the emissive properties of dye-doped PVP nanofibers produced by electrospinning. Journal of Physics and Chemistry of Solids 2014；75：1365-71.
[33] 黃效民. 黑色素之功能. 食品工業月刊, 四月號 1996：8-12.
[34] Iwata M, Corn T, Iwata S, Everett MA, Fuller BB. The relationship between tyrosinase activity and skin color in human foreskins. Journal of investigative dermatology 1990；95：9-15.
[35] Iozumi K, Hoganson GE, Pennella R, Everett MA, Fuller BB. Role of tyrosinase as the determinant of pigmentation in cultured human melanocytes. Journal of Investigative Dermatology 1993；100：806-11.
[36] Pomerantz S, Ances I. Tyrosinase activity in human skin. Influence of race and age in newborns. Journal of Clinical Investigation 1975；55：1127.
[37] Anton F. Artificial thread and method of producing same. Google Patents； 1940.
[38] Formhals A. Method and apparatus for spinning. US patent 1939；2160962.
[39] Liu Y, Zhang L, Sun X-F, Liu J, Fan J, Huang D-W. Multi-jet electrospinning via auxiliary electrode. Materials Letters 2015；141：153-6.
[40] Kim IG, Lee J-H, Unnithan AR, Park C-H, Kim CS. A comprehensive electric field analysis of cylinder-type multi-nozzle electrospinning system for mass production of nanofibers. Journal of Industrial and Engineering Chemistry 2015；31：251-6.
[41] 林孟辰. 聚乙烯醇/海藻酸鈉/白芷複合敷料之製備技術與特性評估. 逢甲大學 2014.
[42] Bognitzki M, Czado W, Frese T, Schaper A, Hellwig M, Steinhart M, et al. Nanostructured fibers via electrospinning. Advanced Materials 2001；13：70-2.
[43] 焦晓宁, 郭光海. 水针压力对水刺非织造布质量的影响. 天津工业大学学报 2003；22：30-2.
[44] Wang K-H, Lin R-D, Hsu F-L, Huang Y-H, Chang H-C, Huang C-Y, et al. Cosmetic applications of selected traditional Chinese herbal medicines. Journal of ethnopharmacology 2006；106：353-9.
[45] Lam RY, Lin Z-X, Sviderskaya E, Cheng CH. Application of a combined sulphorhodamine B and melanin assay to the evaluation of Chinese medicines and their constituent compounds for hyperpigmentation treatment. Journal of ethnopharmacology 2010；132：274-9.
[46] Cooley JF. Apparatus for electrically dispersing fluids. Google Patents； 1902.
[47] Baumgarten PK. Electrostatic spinning of acrylic microfibers. Journal of colloid and interface science 1971；36：71-9.
[48] Lin T, Fang J, Wang H, Cheng T, Wang X. Using chitosan as a thickener for electrospinning dilute PVA solutions to improve fibre uniformity. Nanotechnology 2006；17：3718.
[49] 楊冬芝. 殼聚醣/聚乙烯醇的電紡絲及其交聯研究. 中國化工學會2006.
[50] Hu Z-M, Zhou Q, Lei T-C, Ding S-F, Xu S-Z. Effects of hydroquinone and its glucoside derivatives on melanogenesis and antioxidation： Biosafety as skin whitening agents. Journal of dermatological science 2009；55：179-84.
[51] Ye Y, Chou G-X, Mu D-D, Wang H, Chu J-H, Leung AK-M, et al. Screening of Chinese herbal medicines for antityrosinase activity in a cell free system and B16 cells. Journal of ethnopharmacology 2010；129：387-90.
[52] Enayati M, Ahmad Z, Stride E, Edirisinghe M. Size mapping of electric field-assisted production of polycaprolactone particles. Journal of The Royal Society Interface 2010：rsif20100099.
[53] Machmudah S, Kanda H, Okubayashi S, Goto M. Formation of PVP hollow fibers by electrospinning in one-step process at sub and supercritical CO 2. Chemical Engineering and Processing： Process Intensification 2014；77：1-6.
[54] Mi S, Kong B, Wu Z, Sun W, Xu Y, Su X. A novel electrospinning setup for the fabrication of thickness-controllable 3D scaffolds with an ordered nanofibrous structure. Materials Letters 2015；160：343-6.
[55] Salles THC, Lombello CB, d'Ávila MA. Electrospinning of Gelatin/Poly (Vinyl Pyrrolidone) Blends from Water/Acetic Acid Solutions. Materials Research 2015；18：509-18.
[56] Zhang Z, Wu Y, Wang Z, Zou X, Zhao Y, Sun L. Fabrication of silver nanoparticles embedded into polyvinyl alcohol (Ag/PVA) composite nanofibrous films through electrospinning for antibacterial and surface-enhanced Raman scattering (SERS) activities. Materials Science and Engineering： C 2016；69：462-9.
[57] 楊怡寬. 以靜電紡絲方法備製聚丙烯腈/奈米碳管複合材料之研究. 東海大學 2014.
[58] 汪建民. 材料分析： 中國材料科學學會發行； 1998.
[59] http：//104lab.blogspot.tw/2011/12/blog-post_9503.html.
[60] http：//www.aitmy.com/news/201310/07/news_53896.html.
[61] Yuan Y, Lee TR. Contact angle and wetting properties. Surface science techniques： Springer； 2013. p. 3-34.
[62] Ling LL, Schneider T, Peoples AJ, Spoering AL, Engels I, Conlon BP, et al. A new antibiotic kills pathogens without detectable resistance. Nature 2015；517：455-9.
[63] Kenawy E-R, Bowlin GL, Mansfield K, Layman J, Simpson DG, Sanders EH, et al. Release of tetracycline hydrochloride from electrospun poly (ethylene-co-vinylacetate), poly (lactic acid), and a blend. Journal of controlled release 2002；81：57-64.
[64] 林怡沛. 台灣蘋果及雞血藤之抗氧化活性成分探討. 臺北醫學大學生藥學研究所學位論文 2005：1-192.
[65] 吳宥霖. 桑白皮萃取物製劑之效能與物理性能研究. 大仁科技大學製藥科技研究所碩士論文台中 , 台灣 2009.
[66] 林厚文, 韩公羽. 中药玉竹有效成分研究. 药学学报 1994；29：215-22.
[67] Fernandes J, Correia DM, Botelho G, Padrão J, Dourado F, Ribeiro C, et al. PHB-PEO electrospun fiber membranes containing chlorhexidine for drug delivery applications. Polymer Testing 2014；34：64-71.
[68] Nirmala R, Kalpana D, Jeong JW, Oh HJ, Lee J-H, Navamathavan R, et al. Multifunctional baicalein blended poly (vinyl alcohol) composite nanofibers via electrospinning. Colloids and Surfaces A： Physicochemical and Engineering Aspects 2011；384：605-11.
[69] Fong H, Chun I, Reneker D. Beaded nanofibers formed during electrospinning. Polymer 1999；40：4585-92.
[70] 行政院衛生署藥食品檢驗局，中藥檢驗方法專輯(十). 1996：224-357頁.
[71] Lamke L-O, Nilsson G, Reithner H. The evaporative water loss from burns and the water-vapour permeability of grafts and artificial membranes used in the treatment of burns. Burns 1977；3：159-65.
[72] Ruiz-Cardona L, Sanzgiri YD, Benedetti L, Stella VJ, Topp EM. Application of benzyl hyaluronate membranes as potential wound dressings： evaluation of water vapour and gas permeabilities. Biomaterials 1996；17：1639-43.
[73] Li Y. The science of clothing comfort. Textile progress 2001；31：1-135.