HPLC法测定不同来源重楼药材中7种甾体皂苷的含量

Determination of 7 kinds of steroidal saponins in Rhizoma Paridis from different sources by HPLC

目的分析不同来源重楼属植物的主要活性成分,比较7种甾体皂苷的含量,为重楼药材的鉴别和品质评价提供参考依据。方法采用HPLC法测定不同来源重楼属15批药材中重楼皂苷Ⅶ、重楼皂苷H、重楼皂苷Ⅵ、重楼皂苷Ⅱ、薯蓣皂苷、重楼皂苷Ⅰ和重楼皂苷Ⅴ的含量。色谱条件为Inertsil ODS-3C18色谱柱(150mm×4.6mm,5μm);流动相为乙腈-水(50∶50);检测波长为203nm;流速为1.0mL·min^(-1);柱温为35℃;进样量为20.0μL。结果 15批样品中,重楼皂苷Ⅶ、重楼皂苷Ⅵ、重楼皂苷Ⅱ和重楼皂苷Ⅰ4种皂苷含量之和均在1.849~20.222mg·g^(-1)之间;样品中7种皂苷含量之和在2.351~37.366mg·g^(-1)之间。精密度、稳定性和重复性实验的RSD值均≤3.0%,7种皂苷的平均加样回收率均在95.92%~103.20%之间(RSD≤3.0%,n=5)。结论因产地或种类不同,重楼皂苷的含量存在一定差异。基于15批重楼样品的HPLC图谱,构建了重楼药材的HPLC指纹图谱,实验结果可用于重楼的真伪鉴别和质量评价。

Objective To study the principal active components and to determine the 7kinds of steroidal saponins of Rhizoma Paridis plants from different sources and establish a reference for the identification and quality evaluation.Methods Polyphyllin Ⅶ,polyphyllin H,polyphyllinⅥ,polyphyllinⅡ,polyphyllinⅠ,polyphyllin Ⅴ and dioscin of 15 batches of Rhizoma Paridis plants from different sources were analyzed by HPLC on an Inertsil ODS-3C18（150mm×4.6mm,5μm）column.The mobile phase was composed of acetonitrile-water（50∶50）at a flow rate of 1.0mL·min^-1.The column temperature was 35 ℃.Accordingly,the detection wavelength was set at 203 nm and the injection volume was 20.0μL.Results In the samples,the total content of polyphyllin Ⅶ,polyphyllin Ⅵ,polyphyllin Ⅱ and polyphyllin Ⅰ was between 1.849-20.222 mg·g^-1,and the total content of polyphyllin Ⅶ,polyphyllin H,polyphyllinⅥ,polyphyllinⅡ,polyphyllinⅠ,polyphyllinⅤand dioscin was between 2.351-37.366mg·g^-1.The results showed this method had high precision,good stability and repeatability（RSD≤3.0%）,and the average recoveries were 95.92%-103.20%（RSD≤3.0%,n=5）.Conclusion The results showed that the difference of polyphyllins content of Rhizoma Paridis plants varied from different producing areas and variety classes.HPLC fingerprints of the 15 batches of Rhizoma Paridis plants were established,and could be used for the distinguishing the authenticity,controlling and determing the Rhizoma Paridis.