化妆品美白原料筛选的体外评价体系建立与应用

Establishment and Application of am in Vitro Evaluation System forSelection of Cosmetic Whitening Materials

目的:基于黑色素细胞(B16-F10)建立一种简便、有效的化妆品美白功效原料的体外筛选体系。方法:裂解B16-F10细胞提取细胞酪氨酸酶,探究粗酪氨酸酶提取液的酶活力,构建基本反应体系,优化反应体积、反应比例和反应时间等条件,基于此反应体系评估化妆品原料的潜在美白功效。研究化妆品原料对B16-F10细胞活力影响,采用光学显微镜拍照法检测细胞分泌黑色素的含量。结果:细胞酪氨酸酶活力约为31.70 U/mg。筛选体系的反应条件为总反应体系体积100μL、底物左旋多巴体积20μL、质量浓度1 mg/mL、细胞酪氨酸酶提取液体积30μL、反应时间80 min。光学分析法研究细胞内黑色素合成及分泌的含量,验证了原料光甘草定和传明酸的美白功效。结论:基于黑色素细胞,建立了一种有效的化妆品原料美白功效的体外筛选体系。

Objective:Based on melanocytes(B16-F10),we established a simple and effective in vitro screening system for cosmetic whitening ingredients.Method:First,we cracked B16-F10 cells to extract cytotyrosinase,then explored the enzyme activity of crude tyrosinase extract,constructed a basic reaction system,optimized the reaction volume,reaction proportion and reaction time,and evaluated the potential whitening efficacy of cosmetic raw materials based on this reaction system.To study the effect of cosmetics raw materials on the viability of B16-F10 cells,we used optical microscopy to detect the content of melanin secreted by the cells.Results:The activity of tyrosinase was 31.70 U/mg.The reaction conditions of the screening system were as follows:total reaction system volume 100μL,substrate volume 20μL,concentration 1 mg/mL,cell tyrosinase extract liquid volume 30μL,reaction time 80 min.The content of melanin synthesis and secretion in cells was studied by optical analysis,and the whitening effect of glycyrrhizin and tranamine acid was verified.Conclusion:Based on melanocytes,an effective in vitro screening system for whitening efficacy of cosmetic raw materials was established.