丹参茎叶酚酮有效部位的提取纯化工艺研究

Optimization of extraction and purification technology for phenolic acids and flavonoids in stems and leaves of Salvia miltiorrhiza

目的对丹参茎叶酚酮有效部位提取纯化工艺进行优选。方法采用超高效液相色谱仪检测,以丹参茎叶中酚酸类(丹参素、原儿茶酸、原儿茶醛、咖啡酸、丹酚酸B、迷迭香酸、紫草酸)及黄酮类成分(芦丁、异槲皮苷、山柰酚-3-O-芸香糖苷、紫云英苷)二者提取量、总提取量及浸膏得率为评价指标,通过单因素及正交试验考察提取方法、提取溶剂、料液比、提取时间及提取次数对丹参茎叶提取工艺的影响;采用大孔吸附树脂纯化丹参茎叶提取物并对纯化工艺参数进行考察,确定最佳纯化工艺。结果以50%乙醇8倍量回流提取3次,每次提取1.0 h为最佳提取工艺;以AB-8型大孔吸附树脂纯化,1.0 g/m L药液上样,上样量为每10克干树脂上样1.5 g干燥提取物,40%乙醇洗脱,洗脱用量3 BV为最佳纯化工艺,酚酸类及黄酮类成分总纯度可达到41.83%。结论优选的提取工艺稳定可行,适用于丹参茎叶酚酮有效部位的提取纯化,可为丹参茎叶的进一步开发提供参考。

Objective To study the optimum condition for the extraction and purification of phenolic acids and flavonoids in the stems and leaves of Salvia miltiorrhiza. Methods The contents of phenolic acids and flavonoids were determined by ultra performance liquid chromatography（UPLC）. The extraction process was evaluated by single factor test and orthogonal design with yield of dry extract and the content of phenolic acids（including danshensu, protocatechuic acid, protocatechualdehyde, caffeic acid, salvianolic acid B, rosmarinic acid, and lithospermic acid） and flavonoids（including rutin, isoquercitrin, kaempferol-3-O-rutinoside, and astragalin） as index. The effects of extraction method, extraction solvent, ratio of material to liquid, extracting time, and extracting times on the extraction of stems and leaves of S. miltiorrhiza were investigated. Macroporous adsorption resin was used to purify the sample, and the purification process parameters were investigated to determine the optimum purification process. Results The optimum condition for the extraction of the stems and leaves of S. miltiorrhiza is that eight times of 50% ethanol for three times reflux extraction and 1 h for each time and AB-8 macroreticular resin was selected for the purification. Optimum process was as following： The concentration of sample solution was 1.0 g/m L; The loading quantity of the sample was 0.15 g dried extract of per gram; The resin column chromatography was eluted with 3 BV of 40% ethanol. Under these conditions, the total purity of phenolic acids and flavonoids could reach 40.83%. Conclusion The optimum technology is stable and feasible for the extraction and purification of phenolic acids and flavonoids in the stems and leaves of S. miltiorrhiza, and can provide reference for further development and utilization.