[关键词]
[摘要]
目的 借助UHPLC-Q-Exactive Orbitrap HRMS技术快速鉴定补中益气汤的化学成分,以及小鼠ig后主要入血、组织移行成分分布特征。方法 取12只健康雄性C57/BL6J小鼠,3只空白组ig纯化水,9只给药组按21 g/kgig补中益气汤提取液,分别在给药后0.5、1、3 h各处理3只,收集血清和组织样本,前处理采用蛋白沉淀法。采用ACQUITY UPLC BEH C18色谱柱(100 mm×2.1 mm,1.7 µm),以0.1%甲酸水溶液(A)-甲醇(B)为流动相,梯度洗脱分离。质谱采用正、负离子一级全扫描/数据依赖二级扫描模式采集数据,综合离子峰相对保留时间、一级碎片离子信息,Xcalibur 4.2软件拟合组成元素。通过对照品、文献以及数据库提供的二级碎片离子信息比对,实现对补中益气汤化学成分及入血、各组织原型移行成分的鉴定。结果 从补中益气汤中共鉴定出286种化学成分,包括黄酮类117种、萜类109种、有机酸类29种、苯酞类13种、香豆素5种以及其他类13种(苯乙醇苷类2种、苯丙醇苷类1种、色原酮类2种、色酮类1种、生物碱类1种及木脂素类1种)。在原方鉴定成分的基础上对小鼠ig补中益气汤后取样0.5、1、3 h检测到血清中101、100、102种;肺脏146种、心脏88种、肝脏149种、脾脏138种、肾脏118种、回肠205种;胫骨前肌107种和腓肠肌116种原型移行成分。结论 该研究利用高分辨质谱技术完成了补中益气汤化学物质组及入血、组织移行成分的快速鉴定,可为补中益气汤效应物质的解析及药效成分在不同组织中的药动学研究提供参考。
[Key word]
[Abstract]
Objective To rapidly identify the chemical components of Buzhong Yiqi Decoction and the distribution characteristics of the main absorbed and tissue-migrating constituents in mice after intragastric administration by UHPLC-Q-Exactive Orbitrap HRMS. Methods A total of 12 male C57/BL6J mice were taken, divided into blank group (n = 3) and treatment group (n = 9), the blank group was gavaged with pure water, the treatment group was gavaged with Buzhong Yiqi Decoction extract at 21 g/kg. Three mice were sacrificed at 0.5, 1 and 3 h after the administration of the drug, respectively, and serum and tissue samples were collected, and the pretreatment process was performed by protein precipitation. Using an ACQUITY UPLC BEH C18 column (100 mm×2.1 mm, 1.7 µm), the mobile phase was 0.1% formic acid aqueous solution (A)-methanol (B) with gradient elution. Mass spectrometry collected data using a first-stage full scan of positive and negative ions/data-dependent second-stage scan mode, and based on the relative retention time of ion peaks and the information of primary fragment ions, the constituent elements were fitted with Xcalibur 4.2 software. Through the comparison of reference substances, literature and secondary fragment ion information provided by the database, the identification of the chemical components of Buzhong Yiqi Decoction and the components that enter the blood and migrate to various tissues was achieved. Results A total of 286 chemical constituents were identified from Buzhong Yiqi Decoction, 117 kinds of flavonoids, 109 kinds of terpenes, 29 kinds of organic acids, 13 kinds of phthalein, 5 kinds of coumarins and 13 other kinds of compounds (two kinds of phenylethanol glycosides, one kind of phenylpropanol glycoside, two kinds of chromones, one kind of chromone, one kind of alkaloid and one kind of lignan). On the basis of identified compounds in the extract, 101, 100, and 102 kinds of compounds were detected in the 0.5, 1, and 3 h of mice serum smples after intragastric administration of Buzhong Yiqi Decoction. A total of 146, 88, 149, 138, 118, 205, 107 and 116 kinds of prototype compounds were identified in lung, heart, liver, spleen, kidneys, ileum, anterior tibialis and gastrocnemius, respectively. Conclusion In this study, the chemical components of Buzhong Yiqi Decoction and the absorbed components into serum and tissue-migrating components were rapidly identified by HRMS, which could provide a reference for the analysis of the effective substances of Buzhong Yiqi Decoction and the in-depth study of the pharmacokinetics of the effective components in different tissues.
[中图分类号]
R284.1
[基金项目]
上海市进一步加快中医药传承创新发展三年行动计划资助[ZY(2025-2027)-3-1-1];国家中医药管理局中药创新能力提升项目(BJTH 2024-02F-0009);上海市卫健委医学新技术研究与转化种子计划项目(2024ZZ1006);上海市教委中成药临床评价平台项目(A1-U2 205-0103);上海市科委科技启明星项目(24QA2709200);上海市东方英才计划青年项目(QNWS2024054);上海市自然基金项目(24ZR1467100)