超高效液相色谱与飞行时间质谱联用技术鉴定蒙药蓝盆花在大鼠体内效应成分及其代谢物

Identification of the Metabolites and Effective Components of Flos scabiosae Extract in Rats Using UHPLCQ-Tof-MS / MS Method

目的 研究蒙药蓝盆花在大鼠体内的主要效应成分及其代谢产物。方法 采用Welch Ultimate UHPLC C18柱（2.1mm×100 mm,1.7μm）;柱温：25℃;流动相为乙腈-0.1%甲酸水,梯度洗脱;流速：0.35 m L·min-1。进样量：5μL。SD大鼠灌胃蓝盆花提取物后,收集0-24 h尿液,0-24 h粪便和0-12 h血浆制备含药样品,并与相同条件下空白生物样品及药材提取物图谱对比,采用Peak View v1.2数据处理软件,通过分析化合物保留时间、精确分子质量、元素组成、同位素丰度和一级、二级质谱图,筛选和鉴定尿液、粪便、血浆中的蓝盆花成分及其代谢物。结果 建立了大鼠灌胃蓝盆花提取物后尿液、粪便和血浆中蓝盆花活性物质及其代谢物的鉴定方法,共鉴定出28个效应成分和31个代谢产物。结论 蓝盆花黄酮类化合物代谢主要途径包括氧化、硫酸化、甲基化、葡萄糖醛酸化等。本实验所建方法简便高效,为蓝盆花药效物质基础的阐明提供了依据。

OBJECTIVE To identify the metabolites of Flos scabiosae extract in rats after oral administration for exploring its metabolism mode in vivo. METHODS Welch Ultimate UHPLC Cls column （2. 1 mm x 100 mm, 1.7 μm） was used for the a- nalysis, and the column temperature was maintained at 25 ℃. Gradient elution was conducted with mobile phase consisting of 0. 1% acetic acid water （A） and acetonitrile （B） at a flowing rate of 0.35 mL · min -1. The injection volume was 6 μL. Urine, plasma, and feces samples were collected during 0 - 24 h or 0 - 12 h after oral administration of Flos scabiosae extract to rats. The samples and control samples were analyzed by UHPLC-Q-T MS/MS in the scanning mode to acquire the full-scan chromatograms. The data acquisition and processing analysis were conducted using compound retention time, precise molecular mass, elemental composition, isotopic abundance, and mass spectra with Peak View vl. 2 data processing software. Metabolites were identified by comparison of chromatograms of the urine, feces and plasma samples after administraion of Flos scabiosae extract with those of the blank biological samples and the herb extract. RESULTS The developed method is applicable to the analysis and identification of metabolites of Flos scabiosae extract in biological matrices after oral administration. Based on the investigation of Flos scabiosae extract in rats, 28 parent compounds with 31 metabolites were detected in rat urine, feces, and plasma. CONCLUSION In this study, the developed method is simple and efficient. The main metabolism pathways of Flos scabiosae extract in rats may in- clude oxidation, demethylation, glucuronidation, and sulfation. This study provides a novel pattern and illumination for the re- search of material basis of Flos scabiosae.