Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China; University of Chinese Academy of Sciences, 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 Sciences, Shanghai, 201203, China.
J Pharm Biomed Anal. 2021 May 10;198:113989. doi: 10.1016/j.jpba.2021.113989. Epub 2021 Feb 26.
In vivo metabolite profiling of herbal medicines remains a challenge due to the complex chemical composition and drastic interference from biological matrix. In this study, a systematic strategy was established for comprehensive metabolite profiling of Danqi Tongmai (DQTM) tablet, a combination of salvianolic acids and notoginsenosides, in rats after oral administration. This strategy was composed of six steps. Firstly, the rat plasma and tissue samples were collected at multiple time points to increase the representativeness of samples. Secondly, different sample preparation methods were systematically investigated including protein precipitation, liquid-liquid extraction and solid-phase extraction to obtain superior extraction efficiency for both salvianolic acids and notoginsenosides. Thirdly, the MS acquisition method was optimized by splitting the full scan range into two separate segments to improve the detection capability for minor components. Fourthly, an extended polygonal mass defect filter (EP-MDF) model was constructed to filter potential metabolites of salvianolic acids and notoginsenosides, and remove large amounts of interference ions. Fifthly, ion intensity-based time point-staggered precursor ion list (IITPS-PIL) was generated to trigger more targeted MS/MS acquisition for potential metabolites at the highest concentration. Finally, the absorbed prototypes and metabolites were comprehensively characterized by reference standards and MS/MS fragmentation. The proposed strategy significantly improved the detection ability for trace prototypes and metabolites in vivo. A total of 370 components, including 94 prototypes (38 confirmed with reference standards) and 276 metabolites, were tentatively characterized in rat plasma and tissue samples after oral administration of DQTM. Collectively, this paper provided an applicable reference for comprehensive metabolite profiling of herbal medicines in complex biological samples.
由于中草药的复杂化学成分和生物基质的强烈干扰，其体内代谢产物谱分析仍然是一个挑战。在本研究中，建立了一种系统的策略，用于口服丹芪通络片（DQTM）后大鼠体内综合代谢产物谱分析。该策略由六个步骤组成。首先，在多个时间点收集大鼠血浆和组织样品，以增加样品的代表性。其次，系统研究了不同的样品制备方法，包括蛋白质沉淀、液-液萃取和固相萃取，以获得对丹酚酸和三七总皂苷均具有优异提取效率的方法。第三，通过将全扫描范围分为两个单独的片段，优化 MS 采集方法，以提高对微量成分的检测能力。第四，构建了扩展多边形质量缺陷滤波器（EP-MDF）模型，用于筛选丹酚酸和三七总皂苷的潜在代谢物，并去除大量干扰离子。第五，基于离子强度的时间点交错前体离子列表（IITPS-PIL）生成，以在最高浓度时触发对潜在代谢物的更有针对性的 MS/MS 采集。最后，通过参考标准和 MS/MS 碎片对吸收的原型和代谢物进行综合表征。所提出的策略显著提高了体内微量原型和代谢物的检测能力。通过口服 DQTM 后大鼠血浆和组织样品，共鉴定出 370 种成分，包括 94 种原型（38 种用对照品确证）和 276 种代谢物。总之，本文为复杂生物样品中中草药的综合代谢产物谱分析提供了一种可行的参考。
