复方中药黄酮类成分提取工艺优化及抑制黑素合成活性研究

Study on Extractive Technology of Total Flavones from Compound Chinese Herbal Medicine and Its Inhibitory Effect on Melanin Synthesis

目的:以总黄酮为指标,对由当归、丹参、黄芪、黄芩、甘草、白薇六味中草药形成的组方进行提取工艺优化,并通过体外检测对经过UVB刺激后B16黑素瘤细胞黑素合成抑制活性,探究组方提取物美白功能。方法:采用CCK8法检测样品对B16细胞的增殖抑制活性,用Na OH裂解法测定黑素含量,用硝酸铝-亚硝酸钠比色法测定总黄酮含量,以总黄酮为指标,考察提取时间、提取温度、料液比、溶剂4个主要因素,通过单因素和正交试验,研究组方最佳提取工艺。结果:组方最佳提取工艺为提取时间3 h,提取温度85℃,提取料液比1∶20,提取溶剂75%乙醇。通过最佳提取工艺提取所得的组方提取物,对经过UVB刺激后的B16黑素瘤细胞黑素合成均具有显著抑制活性(P<0.01),且呈剂量依赖型。结论:采用最佳提取工艺提取的组方提取率高,具有较佳的美白活性,能为复方美白功能添加剂的开发提供参考。

Objective： The extraction process of Compound Chinese herbal medicine which contains Radix Astragali, Radix Scutellariae, Radix Angelicae Sinensis, Radix Salviae Mihiorrhizae, Radix et Rhizoma Glycyrrhizae and Radix Cynanchi Atrati was optimized with the content of total flavonoids as index. And through the in vitro detection of the inhibitory activities on the melanogenesis of B16 melanoma cells after UVB stimulation, the whitening effect of component herbs extracts was studied. Methods： The cell proliferation inhibitory activity was detected by cck8 assay, the melanin content was detected by NaOH assay, the content of total flavonoids was detected by colorimetry with AI（ NO3 ）3-NaNO2, with the content of total fla- vonoids as index the four major factors of extraction time, extraction temperature, solid-liquid ratio and extraction solvent were investigated. With single factor and orthogonal experiments, the best extraction process of the prescription was explored. Results： The best extraction process of prescription was as follows： extraction time 3 h; extraction temperature 85 ℃ ; extracting solid-liquid ratio 1：20; extraction solvent 75% ethanol. And the prescription extracts showed a significant inhibitory activity （ P 〈 0.01 ） on the melanogenesis of B16 melanoma cells after UVB stimulation in dose-dependent manner. Conclusion： The prescription extracts through the best extraction process has high extraction efficiency and high skin- whitening activity, which can provide a reference for the development of compound whitening efficacy additive.