| 摘要: | 過去的天然物研究中多以單一植物為主，探討其活性與應用價值。然而，單一植物之活性可能無法滿足化粧品開發者之需求，因此本研究擬以複方植物原料作為材料，進行各種活性評估與化粧品應用潛力之探討。因此本研究整合了七種天然物，植物萃取代號分別為A、B、D、L、M、H、I，將新鮮植物以純水為溶劑，經加熱迴流萃取及冷凍乾燥後，所得到之萃取物，進而評估其各項活性，包括細胞存活率、細胞模擬傷口癒合能力、總酚含量、抗氧化能力、質體DNA保護試驗、體外抑制酪胺酸酶活性等試驗，並經篩選後以複方組成進行活性研究，探討其複合性植物原料是否具有活性協同作用，以應用於化粧品之可行性。
結果顯示，以單獨萃取物而言，在細胞存活率部分，於濃度0.1 mg/mL時，3T3纖維母細胞之存活率皆有85%以上。在細胞模擬傷口癒合能力中，七種萃取物於濃度0.1 mg/mL時，以萃取物 H為最佳，細胞癒合率達到70%。在抗氧化活性試驗中，以萃取物B為較佳，DPPH•自由基之清除能力 EC50 (Half maximal of effective concentration) 為7.50 ± 1.11 μg/mL；ABTS+•自由基之清除能力EC50為5.01 ± 0.54 μg/mL；總酚含量為155.36 ± 4.27 mg GAE/g。在質體DNA保護試驗中，於濃度0.1 mg/mL時，所有天然物對質體 DNA皆有保護效果。在體外抑制酪胺酸酶活性試驗中，以萃取物 M較佳，其IC50 (Half maximal of inhibitory concentration)為0.583 ± 0.26 mg/mL。
經篩選後，針對不同的活性試驗，分別組成複方萃取物，再進行活性研究。而在複方萃取物活性試驗的結果顯示，在細胞模擬傷口癒合能力中，組成三種複方 (V1~V3)，以V1為最佳，複方組成以萃取物 H為主，其細胞癒合率達到70%。抗氧化活性試驗中，在DPPH•自由基之清除率試驗，組成六種複方 (W1~W6)，以W5、W6複方為最佳，複方組成以萃取物B為主，其DPPH•自由基之清除率達到60%；ABTS+•自由基之清除率試驗中，組成六種複方 (X1~X6)，以X5、X6複方為最佳，複方組成以萃取物B為主，其ABTS+•自由基之清除率達到50%以上；質體DNA保護試驗中，組成六種複方 (Y1~Y6)，以Y3、Y6對質體DNA具有保護效果，保護率達到70%。在體外抑制酪胺酸酶活性試驗中，組成三種複方 (Z1~Z3)，以Z3為最佳，複方組成以萃取物M為主，其酪胺酸酶抑制率達到18%。
由以上結果，選用了細胞模擬傷口癒合試驗的複方來設計傷口修復產品，並進行產品之穩定性測試。穩定性結果顯示，萃取物複方產品在不同溫度的產品黏度與酸鹼值較單獨萃取物產品的穩定性高，並將此結果應用於傷口修復產品之開發與應用。
In the past, the natural plants research mainly based on single plant, discussion its activity and business value. However, the activity of single plant may not fulfill the cosmetics developer’s requirement. Therefore, this study intends to composite plant as a material, a variety of assessment and cosmetics development. This study integrates the seven kinds of natural plants, plants codes were A, B, D, L, M, H, I. The fresh plant with water as the solvent, dried and frozen after extraction was heated to reflux, and then assessment of extract activity. Including MTT assay, wound healing assay, total phenols test, antioxidant capacity, plasmid DNA protection test, in vitro inhibition of tyrosinase activity and other tests. After screen the extract, composed of composite extracts and study of its activity. Explore the composite extracts whether has synergistic activity for applying to the development of cosmetics.The results showed at the single extract. In the MTT assay, seven extracts at 0.1 mg/mL, the viability of 3T3 fibroblasts had more than 85%. In the wound healing assay, the extract H at 0.1 mg/mL had the best cell migration rate. In the antioxidant capacity, the extract B had the best antioxidant capacity,In the scavenging of radical DPPH (EC50 : 7.50 ± 1.11 μg/mL), trolox equivalent antioxidant capacity (scavenging of radical ABTS+) (EC50 : 5.01 ± 0.54 μg/mL) and total phenols test (155.36 ± 4.27 mg GAE/g). In the plasmid DNA protection test, seven extracts at 0.1 mg/mL had plasmid DNA protective effect. In the in vitro inhibition of tyrosinase activity, the extract M had the best activity (IC50 : 0.583 ± 0.26 mg/mL).After screening of different activity test, seven extracts were composed of composite extracts, and research of its activity. The results showed at the composite extracts. In the wound healing assay, composed of three composite extracts (V1~V3), the composite extracts V1 which mainly composed was extract H had more than 70% cell migration rate. In the antioxidant capacity, composed of six composite extracts (W1~W6) at scavenging of radical DPPH, the composite extracts W5 and W6 which mainly composed was extract B had more than 60% scavenging of radical DPPH. Composed of six composite extracts (X1~X6) at scavenging of radical ABTS+, the composite extracts X5 and X6 which mainly composed was extract B had more than 50% scavenging of radical ABTS+. Composed of six composite extracts (Y1~Y6) at protection of plasmid DNA, the composite extracts Y3 and Y6 which mainly composed was extract B had more than 70% protection of plasmid DNA. Composed of three composite extracts (Z1~Z3) at in vitro inhibition of tyrosinase activity, the composite extracts Z3 which mainly composed was extract M had more than 18% inhibitory of tyrosinase.From the above, chose the composite extracts V1~V3 to design wound repair cosmetic product, and stability testing of the product. Stability testing results had showed, after four weeks, composite extracts product’s viscosity and pH stability at different temperatures was more stability than single extract product. According to the results, composite extracts was able to use in wound repair cosmetic product’s development and application. |
