超快速液相色谱-三重四级杆-线性离子阱质谱法分析人参和红参中皂苷类成分

Comparative Analysis of Ginsenosides in Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra by UFLC-QTRAP-MS/MS

目的建立超快速液相色谱-三重四级杆-线性离子阱质谱(UFLC-QTRAP-MS/MS)同时测定人参及红参药材中原人参二醇型皂苷[人参皂苷Rb_1、Rc、Rb_2、Rd、F_2、20(S)-Rg_3、20(R)-Rg_3、CK]、原人参三醇型皂苷[人参皂苷Re、Rg_1、Rf、20(S)-Rg_2、20(S)-Rh_1、20(R)-Rg_2、20(R)-Rh_1、F_1]、齐墩果烷型皂苷(人参皂苷Ro) 17种目标成分含量的方法。方法采用Synergi^(TM )Hydro-RP 100柱(2. 1 mm×100 mm,2. 5μm),以0. 1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱,柱温40℃,流速0. 4m L·min^(-1),在电喷雾负离子(ESI-)模式下,进行多反应监测(MRM)模式检测;用主成分分析(PCA)、聚类分析(HCA)进行数据处理。结果 17种目标成分在一定浓度范围内均呈良好的线性关系,相关系数大于0. 999 0;精密度、重复性和稳定性良好;加样回收率为96. 69%～102. 01%,RSD值小于5%。PCA与HCA分析结果显示,人参与红参化学成分差异明显,差异显著的成分为人参皂苷20(S)-Rg_3、20(R)-Rg_3、20(S)-Rh_1、20(R)-Rh_1、20(R)-Rg_2。结论该实验为人参及红参药材内在质量的综合评价和全面控制提供新的方法参考,同时可为揭示二者药性、药效差异的物质基础提供依据。

OBJECTIVE To develop a comprehensive analytical method based on UFLC-QTRAP-MS/MS for simultaneous determination of protopanaxadiol [ginsenoside Rb1, Rc, Rb2, Rd, F2, 20(S)-Rg3, 20(R)-Rg3, CK], protopanaxatriol [ginsenoside Re, Rg1, Rf, 20(S)-Rg2, 20(S)-Rh1, 20(R)-Rg2, 20(R)-Rh1, F1] and oleanolic(ginsenoside Ro) in Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra. METHODS Under the optimized chromatographic conditions, good separation for seventeen target compounds was obtained on a SynergiTM Hydro-RP 100 column(2.1 mm×100 mm, 2.5 μm) at 40 ℃ with 0.1% aqueous formic acid (A)/acetonitrile (B) as the mobile phase by gradient elution at a flow rate of 0.4 mL·min-1. The target compounds were analyzed under multiple reaction monitoring (MRM) mode with an ESI source operated in negative ion mode, and principal component analysis (PCA) and hierarchical cluster analysis (HCA)were used for data processing. RESULTS The calibration curves of the 17 components had good linearity (r>0.999 0). The precision, repeatability and stability were all satisfying.The average recoveries of standard addition for the compounds were between 96.69% and 102.01%,and the relative standard deviations were less than 5%. The results of PCA and HCA showed that Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra were clearly distinguished.The main compositions with significant difference were ginsenoside 20(S)-Rg3, 20(R)-Rg3, 20(S)-Rh1, 20(R)-Rh1, and 20(R)-Rg2. CONCLUSION The established method could provide a new technique for the comprehensive evaluation and quality control of Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra, at the same time, it would pave the way for discovering the material basis contributing to the different properties and efficacies of the two medicinal materials.