野木瓜总皂苷提取工艺优化及其美白活性组分筛选

目的:优化野木瓜中总皂苷的提取工艺，并为野木瓜总皂苷活性组分在美白化妆品中的后续开发应用奠定基础。方法:在单因素实验的基础上，以乙醇浓度、液料比、提取温度、提取时间为优化因素，总皂苷得率为评价指标，Box-Behnken法优化野木瓜总皂苷提取工艺；依次用不同极性溶剂对其提取物进行萃取，硅胶柱色谱层析进行分离，获得不同活性组分，MTT法检测B16细胞的增殖抑制能力，多巴氧化法检测酪氨酸酶活性。结果:各因素对野木瓜总皂苷得率影响顺序为:乙醇浓度>提取温度>液料比>提取时间；最佳提取工艺为乙醇浓度79%，液料比25:1 mL/g，提取温度81℃，提取时间89 min，总皂苷平均得率为2.16%；各萃取部位中正丁醇部位对细胞增殖和酪氨酸酶活性抑制效果最好，效果优于对照品熊果苷（P<0.05），其IC₅₀分别为86.61和69.09 μg/mL；正丁醇部位分离得8个活性组分，其中组分Fr.6对B16细胞增殖和酪氨酸酶活性抑制效果最好，IC₅₀分别为80.93和58.99 μg/mL，且抑制酪氨酸酶活性的效果较分离前的正丁醇部位更好（P<0.05）。结论:本文探讨了野木瓜总皂苷的最佳提取工艺，获得的活性组分Fr.6具有良好的美白活性，在美白化妆品产品开发中具有较好的应用前景。

Optimization of the Extration Technology of Total Saponins from Stauntonia chinensis DC.and the Selection of Its Whitening Active Components

Objective:To optimize the extraction process of total saponins from Stauntonia chinensis DC.and lay a foundation for the further development and application of total saponins in whitening cosmetics.Methods:Based on the single factor experiment,the total saponins yield was optimized by ethanol concentration,liquid to material ratio,extraction temperature and extraction time.The extraction process of total saponins was optimized by Box-Behnken method.The extract of Stauntonia chinensis DC.was extraced by solvents with different polarity,and separated by chromatography to obtain different active components.The proliferation inhibition ability was detected by MTT assay and the tyrosinase activity was detected by Dopa oxidation method.Results:The order of the factors affecting the total saponins was:Ethanol concentration>extraction temperature>liquid to material ratio>extraction time.The optimal conditions were determined to be 79%for ethanol concentration,25∶1 mL/g for liquid to material ratio,81℃for extraction temperature,89 min for extraction time,the average yield of total saponins was 2.16%.The n-butanol site had the best inhibition rate on proliferation of B16 cells and tyrosinase activity,which was better than control arbutin(P<0.05),and IC 50 were 86.61 and 69.09μg/mL.A total of 8 active components were obtained from n-butanol.Fr.6 had the best inhibitory effect on B16 cells and tyrosinase activity,of which the IC 50 were 80.93 and 58.99μg/mL,and inhibition effect of tyrosinase was better than n-butanol site before separation(P<0.05).Conclusion:Response surface methodology was used to determine the optimal extraction process of total saponins and the Fr.6 had good whitening activity.This study shows that the Stauntonia chinensis DC.has a good application prospect in the development of whitening cosmetics.