UPLC-MS/MS法同时测定绞股蓝中16个皂苷类成分的含量  

Determination of 16 saponins in Gynostemmatis Pentaphylli Herba based on UPLC-MS/MS

作　　者：温秀萍[1] 翁艳鸿 陈祖倩 许少华 许文[1,2] 林青青[1] 徐伟[1,2] 林羽[1,2] 陈在敏 WEN Xiu-ping;WENG Yan-hong;CHEN Zu-qian;XU Shao-hua;XU Wen;LIN Qing-qing;XU Wei;LIN Yu;CHEN Zai-min(College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Centre of Biomedical Research&Development,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fujian Institute for Food and Drug Quality Control,Fuzhou 350001,China)

机构地区：[1]福建中医药大学药学院,福州350122 [2]福建中医药大学生物医药研发中心,福州350122 [3]福建省食品药品质量检定研究院,福州350001

出　　处：《药物分析杂志》2022年第2期198-210,共13页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金项目(82104321);2019年医疗服务与保障能力提升补助资金(中医药事业传承与发展部分)“全国中药资源普查项目”(财社[2019]39号);福建省科技厅引导性项目(2018Y0049);福建省教育厅省属高校科研专项资助(JK2017024)。

摘　　要：目的:建立超高效液相色谱串联三重四极杆质谱法(UPLC-MS/MS),在12 min内完成对绞股蓝药材中16个皂苷类成分(人参皂苷CK、F_(2)、Rb_(1)、Rb_(3)、Rd、Rg_(2)、Rg_(3)、Rg_(5)、Rg_(6)、Rh_(4)、Rh_(8),绞股蓝皂苷A、XLVI、XLIX,三七皂苷Fd和越南人参皂苷R_(8))的含量测定。方法:采用UPLC-MS/MS法,正离子多反应监测(MRM)模式进行含量测定。色谱条件:采用Waters CORTECS C_(18)色谱柱(2.1 mm×100 mm,1.6μm),流动相为0.1%甲酸水(A)-乙腈(B),梯度洗脱,流速为0.25 mL·min^(-1),柱温为45℃。结果:在所设定的色谱条件下,12 min内完成对绞股蓝药材中上述16个成分的含量测定,在考察的浓度范围内均呈良好的线性关系(r>0.9976),回收率和RSD分别在96.2%~104.0%和1.5%~2.9%,精密度、重复性、稳定性考察均符合分析要求。含量测定结果发现,16个皂苷中,绞股蓝皂苷XLVI、A、XLIX和人参皂苷Rb_(3)含量较高,含量均值分别达到7.083、5.599、1.657、1.283 mg·g^(-1),且不同产地的绞股蓝药材中皂苷类成分存在较大差异,聚类分析可分为Ⅰ、Ⅱ、Ⅲ3类。统计发现,越南人参皂苷R_(8),人参皂苷Rb_(1)、Rb_(3)、Rd、Rg_(3)、Rg_(5)、Rg_(6)、Rh_(8)、F_(2)、CK,绞股蓝皂苷XLVI和三七皂苷Fd成分含量在类型Ⅱ中高于类型Ⅰ和Ⅲ,而绞股蓝皂苷A与XLIX成分含量在Ⅲ类型中高于类型Ⅰ和Ⅱ。结论:本研究所建立的在12 min内完成对绞股蓝中16个皂苷类成分的UPLC-MS/MS定量分析方法简便、快捷、准确,分离效果较好,可为完善绞股蓝中皂苷类成分的质量标准评价提供依据。Objective:To establish an UPLC-MS/MS method for simultaneous determination of 16 majorcomponents(ginsenoside Rh_(4),ginsenoside Rh_(8),ginsenoside Rg_(6),ginsenoside Rg_(5),ginsenoside F_(2),ginsenoside Rd,ginsenoside Rb3,ginsenoside Rb_(1),ginsenoside C-K,ginsenoside Rg_(3),ginsenoside Rg_(2),gypenoside A,gypenoside XLVI,gypenoside XLIX,notoginsenoside Fd and vina-ginsenoside R_(8))in Gynostemmatis Pentaphylli Herba.Methods:The UPLC-MS/MS assay was performed on a Waters CORTECS C_(18)(2.1 mm×100 mm,1.6μm)column with acetonitrile-water(containing 0.1%formic acid)as mobile phase by gradient elution at a flow rate of 0.25 mL·min^(-1).The column temperature was set at 45℃.MS detection was performed with multiple reaction monitoring(MRM)mode using positive electrospray ionization.Results:Under the optimized chromatographic conditions,good separation of 16 saponins were obtained within 12 min.Good linearities were found in the concentration ranges(r>0.9976),the recoveries and RSDs were in the range of 96.2%-104.0% and 1.5%-2.9%respectively.And the precision,repeatability and stability also met the analysis requirements.The contents of gypenoside XLVI,gypenoside A,gypenoside XLIX and ginsenoside Rb_(3) were higher,with the average contents of 7.083,5.599,1.657 and 1.283 mg·g^(-1),respectively.There were significant differences in the contents of gypenoside in samples from different habitats.Through cluster analysis,they could be divided into three categories.The contents of ginsenoside R_(8),ginsenoside Rb_(1),ginsenoside Rb_(3),gynostemma gynostemma ginsenoside XLVI,ginsenoside Rh_(8),ginsenoside Rd,ginsenoside Fd,ginsenoside Rg_(6),ginsenoside F_(2),ginsenoside Rg_(3),ginsenoside CK and ginsenoside Rg_(5) in typeⅡwere higher than those in typeⅠandⅢ.And the contents of gypenosides A and XLIX in typeⅢwere higher than those in typeⅠandⅡ.Conclusion:It is the first report about simultaneous analysis of 16 saponins in Gynostemmatis Pentaphylli Herba by using UPLC-MS/MS method,which affords highly sensitiv

关 键 词：绞股蓝 含量测定 超高效液相色谱串联三重四极杆质谱 人参皂苷 绞股蓝皂苷 三七皂苷 越南人参皂苷R_(8) 

分 类 号：R917[医药卫生—药物分析学]