摘要
目的:建立东北铁线莲中抗肿瘤活性成分3-O-α-L-吡喃鼠李糖-(1→6)-β-D-吡喃葡萄糖-(1→4)-β-D-吡喃葡萄糖-(1→4)-β-D-吡喃核糖-(1→3)-α-L-吡喃鼠李糖-(1→2)-α-L-吡喃阿拉伯糖齐墩果酸苷(W2)的大孔树脂纯化工艺。方法:采用HPLC测定W2含量,通过对9种大孔树脂的吸附-洗脱能力进行考察,筛选最适宜的大孔树脂型号,对选定的大孔吸附树脂进行静态吸附动力学考察和动态梯度洗脱考察,通过单因素试验考察上样液浓度、吸附流速、除杂溶媒用量和流速等参数对大孔树脂纯化工艺的影响。结果:最佳纯化工艺为上样液质量浓度0.4 g·m L-1,以1.5 BV·h-1的流速过LX-17型大孔树脂柱,加水4 BV和40%乙醇5 BV除杂,除杂流速3 BV·h-1,加80%乙醇1 BV洗脱以替换柱内液体,加80%乙醇6 BV富集W2,洗脱流速1.5 BV·h-1。浸膏中W2质量分数43.14%。结论:LX-17型大孔树脂适用于W2的的分离纯化,优选的工艺稳定可行,可得到纯度相对较高的样品,为该成分的后续药理作用研究提供保障。
Objective： To establish a purification process for W2 [3-O-α-L-Rha-（ 1→6）-β-D-Glu-（ 1→4）-β-D-Glu-（ 1 →4）-β-D-Rib-（ 1 →3）-α-L-Rha-（ 1 →2）-α-L-Ara oleanolic saponin] in Clematis manshurica by macroporous resin. Method： The content of W2 was determined by HPLC,and it was used as index. Type of macroporous adsorption resin was screened by inspecting the adsorption and desorption ability of nine candidate macroporous resins. The static adsorption kinetics and dynamic gradient elution of the best type of macroporous resin was investigated, single factor tests were adopted to optimize purification process. Result：Optimum purification process was as following： the concentration of sample solution of 0. 4 g·m L- 1,adsorbed by LX-17 macroporous resin column with a flow rate of 1. 5 BV·h- 1,washed by 4 BV of water and 5 BV of 40%ethanol for removing impurity with a flow rate of 3 BV·h- 1,taking 1 BV of 80% ethanol to remove the solution in column,and eluted by 6 BV of 80% ethanol for enrichment of W2 with a flow rate of 1. 5 BV·h- 1. Under these conditions,the purity of W2 in dry extract was 43. 14%. Conclusion： LX-17 macroporous resin is suitable for separation and purification of W2. This optimized process is reasonable,stable and feasible,which can obtain W2 sample with high purity.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2016年第15期14-17,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
江西中医药大学重点学科青年教师培养计划项目(2013jzzdxk049)
关键词
东北铁线莲
齐墩果酸
皂苷类
大孔吸附树脂
吸附量
Clematis manshurica
oleanolic acid
saponin
macroporous resin
adsorption capacity
