Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Science, Haike Road 501, Shanghai, 201203, China; University of Chinese Academy of Sciences, No.19A Yuquan Road, Beijing, 100049, China.
Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Science, Haike Road 501, Shanghai, 201203, China.
J Chromatogr A. 2021 Apr 26;1643:462029. doi: 10.1016/j.chroma.2021.462029. Epub 2021 Mar 3.
The inherent complexity of traditional Chinese medicines necessitates the application of multi-dimensional information to accomplish comprehensive profiling and confirmative identification of their chemical components. In this study, we display an enhanced strategy by integrating offline superimposed two-dimensional separation (S-2D-LC) with mass defect filter and diagnostic ion filter to comprehensively characterize the alkaloid composition of Fritillariae Pallidiflorae Bulbus (FPB). The superimposed HILIC × RP and UPCC × RP offline two-dimensional liquid chromatography system was constructed with superior orthogonality (R=0.004 and R=0.001) for chromatographic separation. In total, 70 fractions were collected after the first-dimensional chromatographic separation (HILIC and UPCC) and then analyzed by the second-dimensional reversed phase (RP) liquid chromatography coupled with Q-TOF/MS/MS in FAST DDA acquisition mode. A four-step interpretation strategy combining mass defect filter with diagnostic ion filter was developed to rapidly characterize alkaloids in Fritillaria species. Ultimately, a sum of 529 Fritillaria alkaloids were characterized from two botanical origins of FPB. The integrated strategy is practical to efficiently expose and comprehensively characterize more trace and isomeric components in complex herbal medicines.
中药的内在复杂性需要应用多维信息来全面分析和确证其化学成分。本研究采用离线叠加二维分离（S-2D-LC）与质量亏损过滤和诊断离子过滤相结合的增强策略，全面表征了浙贝母生物碱成分。构建了具有优异正交性（HILIC×RP 和 UPCC×RP 的 R 值分别为 0.004 和 0.001）的叠加 HILIC×RP 和 UPCC×RP 离线二维液相色谱系统，用于色谱分离。在第一次一维色谱分离（HILIC 和 UPCC）后收集了 70 个馏分，然后通过二维反相（RP）液相色谱与 Q-TOF/MS/MS 在 FAST DDA 采集模式下进行分析。开发了一种结合质量亏损过滤和诊断离子过滤的四步解析策略，用于快速表征贝母属植物中的生物碱。最终，从两种浙贝母植物来源中鉴定出了 529 种贝母生物碱。该综合策略实用高效，可有效地揭示和全面表征复杂中草药中更多的痕量和同系物成分。
