超高效液相色谱法测定野生与栽培滇重楼根茎及须根中9种甾体皂苷的含量

Determination of 9 Steroidal Saponins in Rhizome and Fibrous Roots of Wild and Cultivated Paris polyphylla var. yunnanensis by UPLC

目的建立超高效液相色谱(UPLC)法同时测定滇重楼根茎及须根中伪原薯蓣皂苷、重楼皂苷VII、重楼皂苷H、重楼皂苷VI、重楼皂苷II、薯蓣皂苷、纤细薯蓣皂苷、重楼皂苷I、重楼皂苷V等9种甾体皂苷含量的方法,并对32个野生品与栽培品滇重楼的甾体皂苷含量进行评价。方法采用UPLC法,色谱柱为ACQUITY UPLC BEH C18(2.1 mm×150 mm,1.7μm),流动相为乙腈-水,检测波长为203 nm,流速为0.2 mL·min-1,进样体积为5μL,柱温为30℃。结果9种甾体皂苷分离良好,方法学考察均满足分析要求。32个不同产地滇重楼样品甾体皂苷含量差异较大,其中根茎中多数都能检测到9种甾体皂苷,重楼皂苷I、重楼皂苷II含量丰富,重楼皂苷VI、纤细薯蓣皂苷含量相对较低。须根均不含有伪原薯蓣皂苷、薯蓣皂苷、纤细薯蓣皂苷、重楼皂苷V,但重楼皂苷VI、VII、H含量丰富,且含量远高于根茎。主成分分析和聚类分析结果表明,多数产地野生滇重楼与栽培滇重楼根茎及须根中甾体皂苷未能完全分开。独立样本t检验结果表明,除根茎中重楼皂苷VII、须根中重楼皂苷H外,其余活性成分含量均无显著差异,无法区分野生品与栽培品,与聚类分析结果相一致。结论滇重楼栽培品可以代替野生品药用,以保护匮乏的野生滇重楼资源,须根可被保留,用作药物原料。

Objective To establish a method for the simultaneous determination of 9 steroidal saponins in rhizome and fibrous root of Paris polyphylla var.yunnanensis,and to evaluate the content of steroidal saponins in 32 wild and cultivated products of Paris polyphylla var.yunnanensis.Methods Nine steroidal saponins were determined by UPLC on an ACQUITY UPLC BEH C18 column(2.1 mm×150 mm,1.7μm)with a mobile phase consisting of acetonitrile and water.The detection wavelength was 203 nm,the flow rate was 0.2 mL·min-1,the injection volume was 5μL,and the column temperature was 30℃.Results All 9 steroidal saponins could be well separated,and the methodological investigation could meet the analysis requirements.Among the 32 samples from different regions were quite different in contents of steroidal saponins.Nine kinds of steroidal saponins could be detected in the rhizome,among which polyphyllin I and polyphyllin II were abundant,while polyphyllin VI and gracillin were relatively low.There were no pseduoprodioscin,dioscin,gracillin,and polyphyllin V in fibrous roots,but the content of polyphyllin VI,polyphyllin VII and polyphyllin H were rich and much higher than those in rhizome.The results of principal component analysis and cluster analysis showed that steroidal saponins in the wild and cultivated Paris polyphylla from most areas could not be separated completely.The results of independent sample t-test showed that,for wild and cultivated products,there was no significant difference in the active components except for polyphyllin VII in rhizome and polyphyllin H in fibrous roots,which was consistent with the results of cluster analysis.Conclusion The cultivated products of Paris polyphylla var.yunnanensis could replace the wild products to protect the scarce wild resources,and the fibrous roots could be preserved and used as pharmaceutical raw materials.