UHPLC-Q-Exactive Orbitrap HRMS 技术分析肾衰乙方的化学成分与大鼠口服后的组织分布_沈沁

沈沁1,2，李荣胜2，段连香1，曹玉1，杨翊清1，樊叶楠1，翁涛涛1，刘伟2，胡静1

（1. 上海中医药大学附属第七人民医院肾病科，上海 200137；2. 上海中医药大学附属曙光医院药学部，国家中医药管理局中药制剂三级实验室，肝肾疾病病证教育部重点实验室，上海市中医临床重点实验室，上海 201203）
*通信作者 胡静，本科，主任医师，从事肾脏系统疾病研究；刘伟，博士，副研究员，研究方向：中药复方药效物质基础及体内过程

摘要

目的：运用UHPLC-Q-Exactive-Orbitrap HRMS 技术分析鉴定肾衰乙方的化学成分，以及大鼠灌胃后主要入血、组织原型成分分布。方法：采用Acquity UPLC BEH C1 8 色谱柱（100 mm×2.1 mm，1.7 μm），以0.1%甲酸水溶液（A）-甲醇（B）为流动相体系，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（Full MS/dd-MS2 ）采集数据，经Xcalibur 4.2 软件获取色谱保留时间以及准分子离子碎片信息，通过对照品比对、文献参考以及自建数据库等成分检测信息，实现对肾衰乙方化学成分及入血、组织原型成分的鉴定。结果：共鉴定出166 种化学成分，包括黄酮类56 种、皂苷类25 种、蒽醌类24 种、有机（酚）酸类23 种、苯酞类14 种、生物碱和环肽类各7 种、木脂素类和香豆素类各3 种以及其他类4 种。大鼠口服肾衰乙方后分别从血清、心、肝、脾、肺、肾、脑、睾丸中检测到54、53、61、47、71、48、53、42 种原型成分。结论：该研究利用高分辨质谱技术完成了肾衰乙方化学成分及主要入血、组织移行成分的快速鉴定，可为肾衰乙方效应物质的解析及作用机制研究提供参考。
关键词 UHPLC-Q-Exactive Orbitrap HRMS；肾衰乙方；化学成分；入血成分；组织分布
中图分类号 R284.1 文献标志码 A
Chemical profiling and tissue distribution study of Shenshuai Yifang after oral administration in rats by UHPLC-Q-Exactive-Orbitrap HRMS
SHEN Qin1,2, LI Rongsheng2, DUAN Lianxiang1, CAO Yu1, YANG Yiqing1, FAN Yenan1, WENG Taotao1, LIU Wei2, HU Jing1
(1.Department of Nephropathy, The Seventh People’s Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200137, China; 2.Department of Pharmacy, The NATCM Third Grade Laboratory of Traditional Chinese Medicine Preparations, Key Laboratory of Liver and KidneyDiseases (Ministry of Education), Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China )
Abstract OBJECTIVE UHPLC-Q-Exactive-Orbitrap HRMS was used to analyze and identify the chemical components of Shenshuai Yifang, as well as to investigate the distribution of major components in blood and tissues after oral administration in rats. METHODS An Acquity UPLC BEH C18 chromatographic column (100 mm×2.1 mm, 1.7 μm) was used as solid phase, while the mobile was 0.1% formic acid (A) and methanol (B) with gradient elution. The mass spectra was collected using the Full scan of positive and negative ions/data-dependent on the secondary scan function (Full MS/dd-MS2) for data collection. Chromatographic retention time and quasi-molecular ion fragment information were acquired by Xcalibur 4.2 software, while the identification of components was compared with reference standards, literature reference, and self -established database. RESULTS A total of 166 chemical components were identified from Shenshuai Yifang, including 56 kinds of flavonoids, 25 kinds of saponins, 24 kinds of anthraquinones, 23 kinds of organic acids and phenolic acids, 14 kinds of phthalides, 7 kinds of cyclic peptides and alkaloids，3 kinds of lignans and coumarins and 4 kinds of others. After oral administration Shenshuai Yifang to rats, 54, 53, 61, 47, 71, 48, 53, 42 prototype components were detected in serum, heart, liver, spleen, lung, kidney, brain and testicles respectively. CONCLUSION In this study, the chemical components and main blood and tissue distribution components of Shenshuai Yifang, were rapidly identified by high resolution mass spectrometry, which can provide reference for the analysis of effective substances and the explore of action mechanism of Shenshuai Yifang.
Key words UHPLC-Q-Exactive Orbitrap HRMS; Shenshuai Yifang; chemical composition; absorded components; tissue distribution