Jin Miao-Miao, Zhang Wen-Dan, Song Geng-Shen, Xu Yan-Mei, Du Ying-Feng, Guo Wei, Cao Liang, Xu Hui-Jun
Department of Pharmaceutical Analysis, School of Pharmacy, Hebei Medical University, Shijiazhuang 050017, China.
Department of Cosmetics, Hebei Institute for Food and Drug Control, Shijiazhuang 050000, China.
J Chromatogr Sci. 2018 Mar 1;56(3):216-224. doi: 10.1093/chromsci/bmx102.
A tissue-smashing based ultra-rapid extraction coupled with ultra-performance liquid chromatography tandem-mass spectrometry (UPLC-MS/MS) method was developed to determine 10 major triterpenoid saponins from Pulsatilla herbs. Compound 4 was characterized as betulinic acid glycoside 3-O-α-arabinopyranosyl-28-O-β-glucopyranosyl-23-hydroxy with HR-ESI-MS, 1H-NMR and 13C-NMR experiment. The MS spectra result showed that the ionization of compound 4 was more efficient in the positive mode. Meanwhile, the ions at m/z 789.6 and m/z 627.5 were selected as precursor and product ion for the determination, respectively. The chromatographic separation was carried out on a Phenomenex Kinetex C18 column using a gradient mobile phase system composed of 0.1% formic acid both in methanol and water at a flow rate of 0.4 mL/min. The detection was performed by multiple reaction monitoring mode, using electrospray ionization in the positive and negative mode. The total run time was 6 min. The calibration curves possessed good linearity with all coefficients higher than 0.9987. The intra- and interday precisions were no more than 4.9%, and the average recoveries were from 97.6% to 103.4% with RSD <4.7%. Moreover, hierarchical cluster analysis was performed to compare and discriminate the Pulsatilla herbs based on the quantitative data. The hierarchical cluster analysis results demonstrated that Pulsatilla chinensis, Pulsatilla cernua, Pulsatilla dahurica, Pulsatilla turczainovii samples could be easily discriminated from each other based on the contents of triterpenoid saponins and the established method is feasible for quality control of Pulsatilla herbs.
建立了一种基于组织捣碎的超快速提取结合超高效液相色谱串联质谱（UPLC-MS/MS）法，用于测定白头翁药材中的10种主要三萜皂苷。通过高分辨电喷雾电离质谱（HR-ESI-MS）、1H核磁共振（1H-NMR）和13C核磁共振（13C-NMR）实验，将化合物4鉴定为桦木酸糖苷3-O-α-阿拉伯吡喃糖基-28-O-β-葡萄糖吡喃糖基-23-羟基。质谱结果表明，化合物4在正离子模式下的电离效率更高。同时，分别选择m/z 789.6和m/z 627.5处的离子作为测定的母离子和子离子。采用Phenomenex Kinetex C18色谱柱进行色谱分离，流动相为甲醇和水均含0.1%甲酸的梯度体系，流速为0.4 mL/min。采用正、负离子模式的电喷雾电离，通过多反应监测模式进行检测。总运行时间为6分钟。校准曲线具有良好的线性，所有系数均高于0.9987。日内和日间精密度均不超过4.9%，平均回收率为97.6%至103.4%，相对标准偏差（RSD）<4.7%。此外，基于定量数据进行了层次聚类分析，以比较和鉴别白头翁药材。层次聚类分析结果表明，基于三萜皂苷的含量，白头翁、朝鲜白头翁、兴安白头翁、细叶白头翁样品能够很容易地相互区分，所建立的方法对于白头翁药材的质量控制是可行的。
