大孔吸附树脂分离纯化柴胡总皂苷工艺研究

Study on the Refined Process of the Total Bupleurum Saikosaponin by Macroporous Resins

目的以柴胡皂苷a含量为指标,考察柴胡总皂苷的最佳分离纯化工艺。方法建立柴胡皂苷a的HPLC测定法,色谱柱为ODS-C18柱(150×4.6mm,5μm),流动相为甲醇:水(70:30),流速为1mL.min-1,检测波长为210nm。以柴胡皂苷a为指标,比较不同型号大孔树脂对于柴胡总皂苷的分离纯化能力,对吸附流速、洗脱溶剂及用量、洗脱流速进行考察。结果柴胡皂苷a在50～250μg.mL-1的范围内,浓度与其峰面积具有良好的线性关系(r=0.9991),平均加样回收率为99.22%,RSD=1.07%(n=6)。确定D101大孔树脂为柴胡总皂苷的纯化材料,其最佳工艺条件是吸附流速为1BV.h-1,洗脱溶剂为70%乙醇,用量为5BV,洗脱流速为2BV.h-1。柴胡总皂苷精制品中柴胡皂苷a含量可达到3%左右。结论该含量测定方法操作简便、重复性好,可用于测定柴胡总皂苷精制品中柴胡皂苷a的含量。D101大孔树脂分离柴胡总皂苷工艺稳定可行,树脂可重复利用,对于柴胡总皂苷的分离精制具有良好的应用前景。

Objective To explore the best refined process of the total I3upleurum saikosaponin. Methods The content of Saikoside A was assayed by HPLC. ODS-C18 column（150x4.6mm, 5lxm） was used. The mobile phase consisted of methanol-water（30：70）. The flow rate was l mL＇min-1. The detection wavelength was 210nm. To compare refined ability of different macroporous resins by assaying Saikoside A by inspecting absorption flow rate, elution solvent and its usage, elution flow rate. Results The calibration curve was in good linearity within the range of 50~250μ g·mL-1 （r=0.9991）, the average recovery was 99.22% （RSD =1 .07%）. The best conditions were： D101 macroporous resin, the absorption flow rate was 1BV·h-1, the elution solvent was 70% ethanol and its usage was 5BV, the elution flow rate was 2BV ·h-1. The content of Saikoside A in the refined artical was about 3%. Gonclusion The assaying method was simple and had good repetitiveness, could be used to assay Saikoside A in the total Bupleurum saikosaponin refined article, The refined process of the total Bupleurum saikosaponin by D101 macroporous resin was feasible. The macroporous resin was recycling, so it had good applied prospect in the refined process of total Bupleumm saikosaponin.