摘要:
目的 建立盾叶薯蓣提取物中4种皂苷含量的快速测定方法.方法 采用HPLC-MS/MS法,以甘草次酸为对照,采用多级反应监测(MRM)法,同时测定薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷、原薯蓣皂苷的含量.用于定量分析的离子反应分别为m/z 867.5→721.3(薯蓣皂苷)、m/z 1 061.6→915.5(甲基原薯蓣皂苷)、m/z 1 029.3→883.1(伪原薯蓣皂苷)、m/z 1 047.6→901.4(原薯蓣皂苷)、m/z469.3→425.3(内标甘草次酸).结果 采用液质联用法测得,薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷、原薯蓣皂苷质量浓度分别在44~220μg/ml、9~60 μg/ml、3~15 μgl、8.8~66 μg/ml内,线性关系良好,R2分别为0.999 1、0.999 8、0.999 6和0.999 3(n=5),平均加样回收率分别为107.1%、90.6%、88.3%、93.4%.结论 所建立方法快速、简便、准确、重现性好,可用于盾叶薯蓣提取物质量控制.%Objective To establish an HPLC-MS/MS method for simultaneous content determination of four kinds of saponin in the rhizome of Dioscorea zingiberensis C.H.Methods An HPLC-MS/MS method was established using multiple-reaction monitoring (MRM) to quantify dioscin,methylprotodioscin,pseudoprotodioscin,protodioscin.Fragmentation reactions were rm/z 867.5→721.3 (dioscin),m/z 1 061.6→915.5 (methylprotodioscin),m/z 1 029.3→883.1 (pseudoprotodioscin),rm/z 1 047.6→901.4 (protodioscin)and m/z 469.3→,425.3 (internal standard glycyrrhetinic acid) separately.Results The mass concentration and the peak area ratio presented good linearity within 44 ~ 220 μg/ml,9 ~ 60 μg/ml,3 ~ 15 μg/ml,8.8 ~ 66 μg/ml,respectively,R2=0.999 1,0.999 8,0.999 6,0.999 3 (n =5) correspondingly.The average recovery (n =9) was 107.1%,90.6%,88.3%,93.4%,respectively.Conclusion This method is proved to be simple,fast,reproduceable and reliable,which can be used for quality control ofDioscorea zingiberensis rhizome extract.