基于UHPLC-Q-Orbitrap HRMS鉴定经典名方举元煎的化学物质组及小鼠口服后的组织暴露成分

Chemical Profile of the Classic Recipe of Juyuan Decoction and its Tissue Distribution in Mice by UHPLC-Q-Orbitrap-HRMS

关键词:  超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UHPLC-Q-Orbitrap HRMS)  举元煎  经典名方  化学成分  入血成分  组织分布

Key words:  Ultra-high-performance liquid chromatography-quadrupole-orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS)  Juyuan Decoction  classical famous formula  chemical constituents  blood-absorbed components  tissue distribution

摘要:

目的 采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UHPLC-Q-Orbitrap HRMS)技术鉴定举元煎的化学物质组及该方口服后在小鼠血清和组织中的暴露成分。方法 取健康雄性C57/BL6J小鼠(n=12),空白组(n=3)灌胃纯化水,给药组(n=9)按43.2 g.kg-1^灌胃举元煎提取液,分别在给药后0.5 h、1.5 h和3 h收集血清和组织样本,采用蛋白沉淀法前处理。质谱数据采集采用正、负离子一级全扫描/数据依赖二级扫描模式,综合离子峰相对保留时间、精确相对分子量等信息,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质谱裂解碎片信息比对,鉴定举元煎原方的化学成分及其口服后的入血原型成分和组织暴露的原型成分。结果 从举元煎中共鉴定出225种化学成分,包括萜(皂苷)类112种、黄酮类61种、有机酸类27种、生物碱类11种、苯乙醇苷类5种、木脂素4种、香豆素类2种以及其他类3种。小鼠灌胃举元煎后血清52种、肺10种、心30种、肝54种、脾56种、肾118种及回肠44种原型成分。其中5-O-阿魏酰奎尼酸、新甘草苷、7,4"-二羟基黄酮、芒柄花素、升麻素苷、升麻苷 A、24-表-7,8-二羟基升麻醇-3-O-木糖苷成分在血清和各组织中均有暴露。结论 借助UHPLC-Q-Orbitrap HRMS技术完成了举元煎化学物质组及其入血成分和组织暴露成分的快速鉴定,为后续举元煎的质量标准建立和多成分药代动力学等研究提供了关键物质信息。

Abstract:

OBJECTIVE To identify the chemical profile of Juyuan Decoction and the exposed components in serum and tissues of mice after oral administration by using ultra-high performance liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) technology. METHODS A total of 12 healthy male C57/BL6J mice were randomly divided into the blank group (n=3) and the administration group (n=9). The blank group was given purified water by gavage, while the administration group was intragastrically administered with Juyuan Decoction extract at a dose of 43.2 g·kg?1. Serum and tissue samples were collected at 0.5 h, 1.5 h and 3 h after administration, and pretreated by protein precipitation method. Mass spectrometry data were acquired in positive and negative ion modes with full MS/data-dependent MS2 scanning. The elemental composition was fitted by Xcalibur 4.2 software based on the relative retention time and accurate relative molecular weight of ion peaks. The chemical components of Juyuan Decoction, as well as the prototype components absorbed into blood and exposed in tissues after oral administration, were identified by comparing with reference standards, literature reports and secondary mass spectrometry fragmentation information from databases. RESULTS A total of 225 chemical components were identified from Juyuan Decoction, including 112 terpenoid (saponin)-type components, 61 flavonoids, 27 organic acids, 11 alkaloids, 5 phenylethanoid glycosides, 4 lignans, 2 coumarins and 3 other components. After intragastric administration of Juyuan Decoction, 52, 10, 30, 54, 56, 118 and 44 prototype components were detected in mouse serum, lung, heart, liver, spleen, kidney and ileum, respectively. Among them, 5-O-feruloylquinic acid, neoglycyrrhizin, 7,4""-dihydroxyflavone, formononetin, cimicifugoside, cimicifugoside A, and 24-epi-7,8-dihydroxycimigenol-3-O-xyloside were exposed in both serum and all tissues. CONCLUSION UHPLC-Q-Orbitrap HRMS technology enables rapid identification of the chemical profile, blood-absorbed components and tissue-exposed components of Juyuan Decoction, which provides key material basis for the subsequent establishment of quality standards and multi-component pharmacokinetic studies of Juyuan Decoction.