- 學位論文
草莓萃取物面膜功能與生物相容性測試
Strawberry Extract Facial Mask Function and Biocompatibility Test
摘要
在眾多水果中，草莓一直是深受大眾喜愛，鮮食、甜點、飲品都有草莓的身影，因非屬亞熱帶水果，經農民多次品種改良，使台灣草莓品種相當多元。草莓種植成本高，係屬高單價之經濟作物，採收時會有部分草莓被淘汰，其並非因病蟲害所致，而是賣像不佳、甜度不足，遭淘汰的草莓如何能有效再利用，並給予草莓新的價值，而衍生出多樣的再製品。故本研究對市售草莓面膜產品，進行驗證其抗氧化功效、美白功能及生物相容性等三項特性，並做科學與量化的分析，希望建立此產品的功能性定位。本研究在抗氧化分析方面，採用總酚含量測定、總類黃酮含量、DPPH自由基清除、烴基自由基清除等四項進行研究，分析面膜液成分的抗氧化功效與文獻中的鮮果柳丁、鮮果火龍果、鮮果木瓜、鮮果芭樂以及鮮果草莓五種新鮮水果作比較與探討。抗氧化分析結果如下，總酚含量為9.04 mg/mL之沒食子酸當量，含量與一顆鮮果柳丁相近，總類黃酮含量為76.9 μg/mL之蘆丁當量 ，含量與一顆鮮果芭樂相近，DPPH自由基清除率為88.7%，清除效果與一顆鮮果芭樂相近，烴基自由基清除率為52.3%，清除效果與一顆鮮果木瓜相近。面膜液成分檢測結果相比鮮果草莓的總酚含量與總類黃酮含量低，面膜液成分對DPPH自由基之清除效果相對比鮮果草莓好。在美白分析方面，將不同濃度的面膜液與纖維母細胞(L-929)、黑色素腫瘤細胞(B16F10)共培養，透過MTT活性分析法，探討面膜成分對於纖維母細胞與黑色素腫瘤細胞活性的影響，本實驗用控制組與不同濃度面膜液組作比較，纖維母細胞的30 μL面膜液組一至三日細胞活性為103.3 %、98.9%、100.5%，60 μL面膜液組一至三日細胞活性為100.8 %、102.1%、101.4%， 90 μL面膜液組一至三日細胞活性為102.1 %、102.5%、100.9%，120 μL面膜液組一至三日細胞活性為102.4%、102.6%、100.7%，不同濃度的面膜液對於纖維母細胞沒有活性的影響，表示對於纖維母細胞沒有毒性產生；黑色素腫瘤細胞的30 μL面膜液組一至三日細胞活性為98.2 %、75.8%、37.4%，60 μL面膜液組一至三日細胞活性為80 %、66.7 %、33.5 %， 90 μL面膜液組一至三日細胞活性為68.4 %、60.1 %、24.1 %，120 μL面膜液組一至三日細胞活性為55.8 %、53.3 %、24.6%，不同濃度的面膜液對於黑色素腫瘤細胞有活性的影響，表示對於黑色腫瘤細胞中黑色素的產生會有影響，因而導致細胞活性下降，表明面膜成分具有美白的功效。在生物相容性分析方面，石墨烯面膜敷料與面膜成分對於纖維母細胞(L-929)增殖與活性作分析，細胞增殖透過紅血球計數法，來探討細胞數目的差異，本實驗用控制組、面膜組及面膜液組作比較，控制組第一日細胞數12.4萬，七日後增長至89.萬，面膜組第一日細胞數13.萬，七日後增長至92.萬，面膜液組第一日細胞數12.6萬，七日後增長至99.萬，透過計數的結果表明面膜、面膜液組的細胞數目比控制組多，代表石墨烯敷料面膜與面膜成分，不會細胞生長有影響。另細胞活性分析透過MTT活性分析法，來探討細胞活性的差異，面膜組一至三日細胞活性為103 .5 %、102.1 %、100.2 %，面膜液組一至三日細胞活性為103.9 %、104%、100.2 %。透過活性的結果表明面膜、面膜液組的活性比控制組高，石墨烯敷料與面膜成分，代表石墨烯敷料面膜與面膜成分，不會細胞生長有影響。綜上，本研究驗證市售草莓面膜產品之抗氧化功效、美白功能及生物相容性等三項特性，其研究結果是具有抗氧化、美白的功效，並且對纖維母細胞無毒性產生。
並列摘要
Among the many fruits, strawberries have always been loved by the public. Strawberries are found in fresh foods, desserts, and drinks. Because they are not subtropical fruits, they have been improved many times by farmers, making Taiwan's strawberry varieties quite diverse. The cost of strawberry planting is high, and it is a high unit price economic crop. Some strawberries will be eliminated when harvested. This is not caused by diseases and insect pests, but because of insufficient sweetness. How can the eliminated strawberries be effectively reused and give strawberry a new value, and derive a variety of re-products.Therefore, in this study, the facial mask products made of strawberry extracts were tested to verify their three characteristics, namely anti-oxidative effect, whitening function and biocompatibility, and to conduct scientific and quantitative analysis, hoping to establish the functional positioning of this product. In terms of antioxidant analysis, this study used four items of total phenolic content determination, total flavonoid content, DPPH free radical scavenging, and hydrocarbon free radical scavenging. Fruits for comparison and discussion. The total phenol content is 9.04 mg/mL equivalent to gallic acid, which is similar to that of orange, the total flavonoid content is 76.9 μg/mL to the equivalent of rutin, which is similar to that of guava, and the scavenging rate of DPPH free radicals is 88.7%, and the scavenging effect is similar to that of guava Similar, the scavenging rate of hydrocarbon radicals is 52.3%, and the scavenging effect is similar to that of papaya. In terms of whitening analysis, different concentrations of mask liquid were co-cultured with fibroblasts (L-929) and melanoma cells (B16F10), and the effect of mask ingredients on the activity of fibroblasts and melanoma cells was explored by MTT activity assay. In this experiment, the control group was compared with the facial mask group with different concentrations at the three day. The cell viability of fibroblasts in the 30 μL facial mask group was 103.3%, 98.9%, 100.5% , and in the 60 μL facial mask group was 100.8%, 102.1%, 101.4% , and in the 90 μL facial mask group was 102.1%, 102.5%, 100.9% , and in the 120 μL facial mask group was 102.4%, 102.6%, 100.7% . The concentration of facial mask liquid has no effect on the activity of fibroblasts, indicating that there is no toxicity to fibroblasts; the cell viability of melanoma in the 30 μL facial mask group was 98.2%, 75.8%, 37.4%, and in the 60 μL facial mask group was 80%, 66.7%, 33.5%, and in the 90 μL facial mask group was 68.4%, 60.1%, 24.1% , and in the 120 μL facial mask group was 55.8%, 53.3%, 24.6%, different concentrations of mask liquid have an effect on the activity of melanoma cells, which means that it will affect the production of melanin in melanoma cells, resulting in a decrease in cell activity, indicating that the ingredients of the mask have a whitening effect. In terms of biocompatibility analysis, the graphene mask dressing and mask components were analyzed for the proliferation and activity of fibroblasts (L-929). The cell proliferation was analyzed by the red blood cell count method to explore the difference in the number of cells. In this experiment, the control group, Compared with the mask group and the mask liquid group, the number of cells in the control group was 12.4*10^4 cell on the first day, and increased to 89.6*10^4 cell after seven days; the number of cells in the mask group was 13.5*10^4 cell on the first day, and increased to 92.8*10^4 cell after seven days; The count was 12.6*10^4 cell, which increased to 99.2 *10^4 cell after seven days. The results of counting showed that the number of cells in the mask and mask liquid group was more than that in the control group, which means that the graphene dressing mask and mask components will not affect cell growth. In addition, the cell activity analysis used the MTT activity analysis method to explore the difference in cell activity. The cell activity of the mask group was 103.5%, 102.1%, and 100.2% in the first to third day, and the cell activity of the mask liquid group was 103.9%,104%, and100.2% in the first to third day. The results of the cell activity showed that the activity of the mask and mask liquid group was higher than that of the control group, and the graphene dressing and mask components, representing the graphene dressing mask and mask components, would not affect cell activity.In summary, this study verified the three characteristics of strawberry extract mask products, including antioxidant effect, whitening function and biocompatibility.