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(2.1 mm×100 mm，1.7 μm);柱温:25 ℃;流动相为乙腈-0.1%甲酸水，梯度洗脱;流速:0.35 mL·min-1。进样量:5 μL。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PeakView 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。
Abstract
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 C18 column (2.1 mm×100 mm, 1.7 μm) was used for the analysis, 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 v1.2 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 include oxidation, demethylation, glucuronidation, and sulfation. This study provides a novel pattern and illumination for the research of material basis of Flos scabiosae.
关键词
蓝盆花 /
超高效液相色谱与飞行时间质谱联用 /
效应成分 /
代谢产物
{{custom_keyword}} /
Key words
Flos scabiosae /
UHPLC-Q-Tof-MS/MS /
effective component /
metabolite
{{custom_keyword}} /
中图分类号：
R282
{{custom_clc.code}}
({{custom_clc.text}})
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Zhonghuabencao Editing Committee of State TCM Administration. Chinese Materia Medica. Mongolian Drugs(中华本草. 蒙药卷). Shanghai:Shanghai Science and Technollog Press, 2004:385.
[2] LI T E,OUYANG H,YANG L J,et al. Research progress on chemical constituents of plants from Scabiosa L. and their pharmacological activities . Chin J Exp Tradit Med Form(中国实验方剂学杂志),2015,21(18):226-230.
[3] ZHANGY S,LI Y Z,CHEN B,et al. Simultaneous quantification of loureirin A and loureirin B in rat urine, feces, and bile by HPLC-MS/MS method and its application to excretion study. Anal Bioanal Chem,2011, 4(400):1181-1187.
[4] WAN Y G, LI N, ELAINE L L,et al. Rapid identification of new minor chemical constituents from Smilacis Glabrae Rhizoma by combined use of UHPLC-Q-TOF-MS, preparative HPLC and UHPLC-SPE-NMR-MS techniques. Phytochem Anal,2015,6 (26):428-435.
[5] QI L W, CHEN C Y, LI P,et al. Structural characterization and identification of iridoidglycosides, saponins, phenolic acids and flavonoids in flos Lonicerae japonicae by a fast liquid chromatography method with diode-array detection andtime-of-flight mass spectrometry. Rapid Commun Mass Spectrom,2009,19(23):3227-3242.
[6] GUO X F,YUE Y D,TANG F, et al. A comparison of C-glycosidic flavonoid isomers by electrospray ionization quadrupole time-of-flight tandem mass spectrometry in negative and positive ion mode. Int J Mass Spectrom,2013,1(333):59-66.
[7] GROSSO C,VINHOLES J,SILVA L R,et al. Chemical composition and biological screening of Capsella bursa-pastoris. Rev Bras Farmacogn,2011,21(4):635-643.
[8] MIZUNO T,UEHARA A,MIZUTA D,et al. Contribution of anthocyanin-flavone copigmentation to grayed violet flower color of Dutch iris cultivar ‘Tiger′s Eye’ under the presence of carotenoids. Sci Hortic,2015,21(186):201-206.
[9] HUANG M,ZHANG Y,XU S,et al. Identification and quantification of phenolic compounds in Vitex negundo L. var. cannabifolia (Siebold,etZucc. ) Hand. -Mazz. using liquid chromatography combined with quadrupole time-of-flight and triple quadrupole mass spectrometers. J Pharm Biomed Anal,2015,15(108):11-20.
YANG Y,WANG H J,YANG J,et al. Chemical profiling and quantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction, a systematic quality control strategy using ultra high performance liquid chromatography combined with hybrid quadrupole-orbitrap and triple quadrupole mass spectrometers. J Chromatogr A,2013,20(1321):88-99.
JI M,LI S J,MA C M. Chemical constituents of the inflorescence of Scabiosacomosa Fisoh and their antioxideand α-glucosidase Inhibitory activities. Acta Acad Med Neimongol(内蒙古大学学报),2014,43(4):398-403.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
基金
江西省青年科学家培养对象资助项目(20142BCB23022);江西民族传统药现代科技与产业发展协同创新中心开放课题资助项目(JXXT201402009)
{{custom_fund}}
PDF(1238 KB)
