粉萆薢总皂苷提取及总皂苷、薯蓣皂苷含量分析

Extraction of total saponin of dioscoreae and content analysis of total saponins and dioscin

目的建立粉萆薢总皂苷(Total saponin of Dioscorea,TSD)提取工艺,测定TSD和薯蓣皂苷的含量。方法乙醇回流法提取TSD,以薯蓣皂苷元为对照品,高氯酸为显色剂,紫外-可见分光光度法测定TSD含量(测定波长409±1 nm);以薯蓣皂苷为对照品,HPLC法测定其中薯蓣皂苷含量(C18色谱柱4.6 mm×250 mm,5μm,流动相为甲醇和水,梯度洗脱,检测波长为208 nm)。结果 TSD提取工艺为10倍量95%乙醇回流提取3次,每次1 h;等量石油醚脱脂,水饱和正丁醇萃取2次,薯蓣皂苷元线性范围为25.4～152.4μg(r=0.999 2),平均回收率为99.39%(RSD=1.62%)。薯蓣皂苷与其他成分分离良好,峰面积与浓度呈良好的线性关系,线性范围为1.0～8.0μg(r=0.999 4),平均回收率为99.22%(RSD=1.56%)。三批粉萆薢药材提取物中的总皂苷的含量分别为61.74%、58.78%和60.47%,薯蓣皂苷的含量分别为33.9%、33.15%、32.26%。结论该提取方法合理,含量测定方法简便、快速、准确、重复性好,可作为TSD和薯蓣皂苷的定量分析。提取物总皂苷及薯蓣皂苷含量能满足药效学实验的要求。

Objective To establish the extraction method of total saponin of dioscorea,and detect total saponin and dioscin in dioscoreae. Methods The TSD was obtained by ethanol reflux extraction method.Spectrophotometer was used to determine the total saponin with perchloric acid as colorimetric reaction and dioscin as the reference substance and measurement wavelength was 409±1 nm. Take dioscin as the reference substance,high-pressure liquid chromatography was used to detect the concentration of dioscin in it（ ACE EXCEL C18 column,4. 6 mm × 250 mm,5 μm,Methanol-water solution as mobile phase,gradient elution,detection wavelength was 208 nm）.Results The best extraction process was as follows： 95% alcohol（ ten times to the sample）,to reflux for one hour and extracted for 3 times. The organic solvent extraction was extracted by isometric petroleum ether and water saturated n-butanol for twice. The linearity range of diosgenin was from 25.4 to 152.4μg（ r = 0.9992）. The average recovery was 99. 39%（ RSD = 1. 62%）. Diosgenin is well separated from other components,and there is a good linear relationship between peak area and concentration. Linear range of was from 1.0 to 8.0 μg（ r = 0.9994）,with the average recovery of 99.22%（ RSD = 1.56%）.The contents of total saponins of Dioscorea in three batches of herbs were 61.74%,58.78% and 60.47%,respectively,and the contents of dioscin were 33.9%,33. 15% and 32. 26%,respectively. Conclusions The extraction method is reasonable,and the quantitative method is simple,rapid,accurate and reproducible,it can be used to detect the concentration of TSD and dioscin. The contents of total saponin and diosgenin can meet the requirements of pharmacodynamics test.