RP-HPLC测定金刚藤中薯蓣皂苷元的含量

RP-HPLC determination of diosgenin in Smilax china L.

目的 建立金刚藤中薯蓣皂苷元的测定方法。方法 采用RP HPLC测定薯蓣皂苷元的含量 ,色谱柱为LichrospherC18(4 6mm× 2 5 0mm ,5 μm) ,流动相为甲醇 ,检测波长为 2 10nm ,峰面积外标法定量。采用正交设计 ,考察发酵时间 (A)、水解时间 (B)、硫酸浓度 (C) 3个因素对薯蓣皂苷水解的影响。结果 薯蓣皂苷元在 5 0～ 35 0 μg·mL-1内具有良好的线性关系 (r =0 9991,n =7) ,平均回收率为 99 97% ,RSD =0 85 % (n =5 )。采用发酵 72h后用浓度为 5 %的硫酸水解 6h ,可使金刚藤中薯蓣皂苷水解完全。结论 RP HPLC是一个简便、快速、分离效果好、精密度高、准确度好的测定金刚藤中薯蓣皂苷元的方法。

OBJECTIVE: To establish a method for determining diosgenin in Smilax china L. METHODS: A lichrospher C18(4.6 mm × 250 mm, 5 μm) column was used with methand as the mobile phase. The detection wavelength was at 210 nm. An external standardization method was used. Three factors on dioscin hydrolysis, i. e. zymolysis time(A), hydrolysis time (B), the concentration of sulfuric acid(C) were studied by orthogoral design. RESULTS: The linear range of 50-350 μg&middotmL-1(r = 0.999 1, n = 7)and an average recovery of 99.97%, RSD = 0.85% (n = 5)were obtained. The optimum hydrolytic condition included zymolysis for 72 h and hydrolysis for 6 h with 5% sulfuric acid. CONCLUSION: RP-HPLC is a simple, rapid and reliable method for diosgenin determination in Smilax china L. Zymolysis is the most critical factor on the hydrolytic yield of diosgenin.