目的 建立HPLC法同时测定不同产地穿山龙药材中原薯蓣皂苷、原纤细薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷、薯蓣皂苷和纤细薯蓣皂苷。方法 穿山龙药材粉末经50%甲醇超声处理。采用Agilent SB-Aq色谱柱（250 mm×4.6 mm，5 μm）；流动相：乙腈–水，梯度洗脱；检测波长203 nm；体积流量1.0 mL/min；柱温25℃；进样体积20 μL。结果 原薯蓣皂苷、原纤细薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷、薯蓣皂苷和纤细薯蓣皂苷在1.054 0～10.540 0、0.519 0～5.190 0、0.287 0～2.870 0、0.021 2～0.212 0、0.061 4～0.614 0、0.061 4～0.614 0 μg线性关系良好，r均大于0.999 0。平均加样回收率分别为100.19%、100.24%、99.87%、100.92%、99.36%、100.03%，RSD值分别为2.03%、2.51%、2.33%、3.07%、2.77%、3.41%。不同产地的穿山龙药材中原薯蓣皂苷、原纤细薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷、薯蓣皂苷和纤细薯蓣皂苷的质量分数分别为1.587～4.837%、0.260～1.950%、0.477～1.317%、0.037～0.187%、0.801～3.813%、0.088～0.524%。结论 该方法快速、简便、准确，可用于穿山龙药材的质量控制。

Objective To establish an HPLC method for simultaneous determination of protodioscin, protogracillin, methyl protodioscin, pseudoprotodioscin, dioscin, and gracillin in Dioscoreae Nipponicae Rhizoma from various of habitats. Methods The powders of Dioscoreae Nipponicae Rhizoma was ultrasonically extracted by 50% methanol. The separation was carried out on Agilent SB-Aq chromatography column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-water with gradient elution. The detection wavelengths were set at 203 nm. The flow rate was 1.0 mL/min, temperature of column was set at 25℃, and volume of injection was 20 μL. Results The linear ranges of protodioscin, protogracillin, methyl protodioscin, pseudoprotodioscin, dioscin, and gracillin were 1.054 0-10.540 0, 0.519 0-5.190 0, 0.287 0-2.870 0, 0.021 2-0.212 0, 0.061 4-0.614 0, and 0.061 4-0.614 0 μg, respectively. And all r values were above 0.999 0. The average recoveries were 100.19%, 100.24%, 99.87%, 100.92%, 99.36%, and 100.03% with RSD 2.03%, 2.51%, 2.33%, 3.07%, 2.77%, and 3.41%, respectively. The contents of protodioscin, protogracillin, methyl protodioscin, pseudoprotodioscin, dioscin, and gracillin in Dioscoreae Nipponicae Rhizoma from various of habitats were 1.587-4.837%, 0.260-1.950%, 0.477-1.317%, 0.037-0.187%, 0.801-3.813%, 0.088-0.524%, respectively. Conclusion The method is rapid, simple, and accurate, and can be used for quality control of Dioscoreae Nipponicae Rhizoma.

天津市科技支撑计划项目（15ZCZDSY00470）；天津市应用基础与前沿技术研究计划（14JCQNJC13800；15JCYBJC29500）