Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, East Road of North 3rd Ring, Chaoyang District, Beijing 100029, China.
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Avenida da Universidade, Taipa, Macao 999078, China.
Anal Chem. 2021 Feb 2;93(4):2541-2550. doi: 10.1021/acs.analchem.0c04602. Epub 2021 Jan 13.
Direct infusion (DI) has an extraordinary high-throughput advantage. Pseudo-targeted metabolomics (PTM) has been demonstrated integrating the merits of both nontargeted and targeted metabolomics. Herein, we attempted to implant DI into the PTM concept to configure a new strategy allowing shotgun PTM. First, a versatile MS/MSALL program was applied to acquire MS and MS spectra. Second, online energy-resolved MS (online ER-MS) was conducted to obtain breakdown graph as well as optimal collision energy (OCE) for each ion transition paired by precursor ion and the dominant product ion. Third, selected reaction monitoring (SRM) was responsible to output a quantitative dataset with a constant length. Moreover, breakdown graph also served as orthogonal structural evidence when matching MS spectra between DI-MS/MS and an in-house library to strengthen structural annotation confidence. To evaluate and illustrate the utility of the new strategy toward shotgun PTM of medicinal plants, in-depth chemome comparison was conducted within three species, all of which are edible medicinal plants and playing essential roles for turning the deserts into the oases. A total of 185 variables participated in the quantitative measurement program. Each diagnostic ion pair was featured with an OCE. Significant species differences occurred, and echinacoside, acteoside, isoacteoside, 2'-acetyl-acteoside, tubuloside B, mannitol, sucrose, betaine, malate, as well as choline were found to be confirmative chemical markers offering primary contributions toward the species discrimination. After cross-validation with LC-MS/MS, DI-MS/MS fortified with the new strategy is an eligible tool for shotgun PTM, beyond plants.
直接进样（DI）具有极高的高通量优势。伪靶向代谢组学（PTM）结合了非靶向和靶向代谢组学的优点，已得到证实。在此，我们尝试将 DI 植入 PTM 概念中，构建一种新的策略，允许进行 shotgun PTM。首先，采用通用的 MS/MSALL 程序获取 MS 和 MS 谱。其次，进行在线能量分辨 MS（online ER-MS）以获得每个离子跃迁的碎裂图谱以及最佳碰撞能（OCE），该离子跃迁由母离子和主要产物离子对组成。然后，选择反应监测（SRM）负责输出具有恒定长度的定量数据集。此外，碎裂图谱还可用作 DI-MS/MS 与内部库之间匹配 MS 谱时的正交结构证据，以增强结构注释的置信度。为了评估和说明该新策略在药用植物 shotgun PTM 中的应用，在三个物种中进行了深入的化学组比较，这三个物种均为可食用的药用植物，对于将沙漠变成绿洲起着至关重要的作用。共有 185 个变量参与了定量测量程序。每个诊断离子对都具有一个 OCE。观察到显著的物种差异，发现了毛蕊花糖苷、毛蕊花苷、异毛蕊花糖苷、2'-乙酰毛蕊花糖苷、芦丁 B、甘露醇、蔗糖、甜菜碱、苹果酸以及胆碱，它们被认为是确认的化学标志物，为物种鉴别提供了主要依据。与 LC-MS/MS 交叉验证后，DI-MS/MS 与该新策略相结合是一种适用于 shotgun PTM 的工具，不仅限于植物。
