Department of Phytochemistry, Second Military Medical University, Shanghai 200433, PR China.
School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, PR China.
Phytomedicine. 2018 May 15;44:164-172. doi: 10.1016/j.phymed.2018.02.007. Epub 2018 Feb 28.
Panax ginseng (PG) is one of the most valuable and frequently used phytomedicine in Asia. In the current Chinese Pharmacopeia, only three ginsenosides, Rg1, Re and Rb1, were set as standard compounds for the quality evaluation of PG. However, only these three compounds could hardly reflect the quality and therapeutic efficacy of this traditional Chinese medicine (TCM).
Quantification analysis of quality markers (Q-markers) in PG is meaningful for determining the quality of this herbal medicine.
By combining the modes of multiple reaction monitoring (MRM) and single ion monitoring (SIM) of tripe quadrupole mass spectrometry (QqQ-MS) through a mode-switching function, a novel, sensitive and effective LC-MS/MS method has been established to simultaneous quantify fifteen Q-markers in PG in one run time cycle. In order to comprehensively evaluate the quality of ten batches of PG, hierarchical clustering analysis (HCA) and the complete linkage method were conducted on the data of the contents of fifteen ginsenosides.
Thirteen Q-markers, including four pairs of isomers with the same product ions and approximately the same retention times, have been well-separated by MRM. Meanwhile, the other two Q-markers with no fragments have also been quantified by SIM. Chromatographic separation was carried out on a reversed-phase C column by stepwise gradient elution with a mobile phase of water (containing 0.1% formic acid, v/v) and acetonitrile. Good linearity was observed with the correlation coefficients (r) greater than 0.99. The intra- and inter-day precisions as well as repeatability of all of the investigated Q-markers were all no more than 5.91%. The average recoveries varied from 83.06% to 116.42%, with relative standard deviation values (RSDs) less than 6.73%. The total contents of the fifteen ginsenosides in ten batches of PG were in the range of 15.54-24.03 mg/g. The results indicated that the growing region has a significant impact on the contents of ginsenosides in PG.
The proposed approach could be readily utilized as a comprehensive approach for determining the consistency of the quality and therapeutic efficacy of PG, and it might be an example for the selection of Q-marker standards for the Chinese Pharmacopoeia.
人参（PG）是亚洲最有价值和最常用的植物药之一。在中国药典中，仅设定了 Rg1、Re 和 Rb1 这三种人参皂苷作为 PG 质量评价的标准化合物。然而，仅这三种化合物很难反映这种中药的质量和治疗效果。
对 PG 中的质量标志物（Q-标志物）进行定量分析，有助于确定这种草药的质量。
通过三重四极杆质谱（QqQ-MS）的多重反应监测（MRM）和单离子监测（SIM）模式的切换功能，建立了一种新的、灵敏有效的 LC-MS/MS 方法，可在一个运行时间周期内同时定量测定 PG 中的 15 种 Q-标志物。为了全面评价 10 批 PG 的质量，对 15 个人参皂苷含量的数据进行了层次聚类分析（HCA）和完全连接法分析。
通过 MRM 很好地分离了 13 种 Q-标志物，包括 4 对具有相同产物离子和大致相同保留时间的异构体。同时，还通过 SIM 对另外两种没有碎片的 Q-标志物进行了定量。色谱分离在反相 C 柱上进行，采用水（含 0.1%甲酸，v/v）和乙腈的分步梯度洗脱。所有调查的 Q-标志物均具有良好的线性关系，相关系数（r）均大于 0.99。所有 Q-标志物的日内和日间精密度以及重复性均不超过 5.91%。平均回收率在 83.06%至 116.42%之间，相对标准偏差（RSD）值均小于 6.73%。10 批 PG 中 15 种人参皂苷的总含量在 15.54-24.03mg/g 之间。结果表明，生长区域对 PG 中人参皂苷的含量有显著影响。
所提出的方法可用于快速确定 PG 质量和治疗效果的一致性，也可为中国药典选择 Q-标志物标准提供参考。
