Department of Pharmaceutical Analysis, School of Pharmacy, Hebei Medical University, Shijiazhuang, 050017, Hebei, China.
Beijing Youcare Kechuang Pharmaceutical Technology Co., Ltd., Beijing, 100176, China.
Anal Bioanal Chem. 2019 Apr;411(10):2071-2088. doi: 10.1007/s00216-019-01631-6. Epub 2019 Feb 8.
Pulsatilla Adans (PSA) herbs (Ranunculaceae) have been widely used in traditional medicine in China and other countries. However, the authentication and quality control of PSA herbs have always been a challenging task due to their similar morphological characteristics and the diversity of the multiple components that exist in the complicated matrix. Herein, a novel integrated strategy combining UHPLC/Q-Orbitrap-MS techniques with chemometrics analysis is proposed for the discrimination of PSA materials. We developed a comprehensive method integrating a nontargeted bidirectionally screened (NTBDS) MS data set and a targeted extraction peak area analysis for the characterization of triterpenoid saponins of PSA from different species. After that, partial least-squares discriminant analysis (PLS-DA) was performed on the obtained MS data set and the parameter variable importance for the projection (VIP) value and P value were employed to screen the valuable MS features to discriminate PSA from different species. In addition, the receiver operating characteristic (ROC) curve is used to verify the reliability of MS features. Finally, heatmap visualization was employed to clarify the distribution of the identified triterpenoid saponins, and four medicinal species of PSA were successfully differentiated. Additionally, 34 constituents were reported in PSAs for the first time, 81 triterpenoid saponins were identified as differential components, and 12 chemical ingredients were characterized as potential chemical markers to differentiate the four officinal PSA herbs. This is the first time that the differences in different PSA herbs have been observed systematically at the chemical level. The results suggested that using the identified characteristic components as chemical markers to identify different PSA herbs was effective and viable. This method provides promising perspectives in the analysis and identification of the ingredients of Chinese herbal medicines, and the identification of similar herbs from the same species.
白头翁（PSA）草药（毛茛科）在中国和其他国家的传统医学中被广泛应用。然而，由于其相似的形态特征和复杂基质中存在的多种成分的多样性，PSA 草药的鉴定和质量控制一直是一个具有挑战性的任务。在此，提出了一种结合 UHPLC/Q-Orbitrap-MS 技术和化学计量学分析的新型综合策略，用于 PSA 材料的鉴别。我们开发了一种综合方法，将非靶向双向筛选（NTBDS）MS 数据集和靶向提取峰面积分析相结合，用于鉴定不同物种的白头翁三萜皂苷。之后，对获得的 MS 数据集进行偏最小二乘判别分析（PLS-DA），并采用参数变量重要性投影（VIP）值和 P 值筛选有价值的 MS 特征，以区分不同物种的 PSA。此外，还使用接收器操作特性（ROC）曲线验证 MS 特征的可靠性。最后，采用热图可视化方法阐明鉴定的三萜皂苷的分布情况，成功区分了四种药用白头翁。此外，首次报道了 PSA 中的 34 种成分，鉴定了 81 种三萜皂苷作为差异成分，12 种化学成分被表征为潜在的化学标志物，用于区分四种药用 PSA 草药。这是首次系统地从化学水平上观察不同 PSA 草药之间的差异。结果表明，使用鉴定的特征成分作为化学标志物来识别不同的 PSA 草药是有效和可行的。该方法为中药成分的分析和鉴定以及同一种植物的类似草药的鉴定提供了有前景的视角。
