UPLC-Orbitrap HRMS法测定小鼠血液中香豆素及其代谢物

发布时间：2018-03-15 12:54

本文选题：香豆素　切入点：-MOP　出处：《河南师范大学学报(自然科学版)》2017年02期 　论文类型：期刊论文

【摘要】：以香豆素为探针底物,建立了测定小鼠血液中香豆素及其代谢物的超高效液相色谱-轨道阱高分辨质谱(UPLC-Orbitrap HRMS)法.以4-甲基伞形酮为内标,样品用乙腈沉淀蛋白后,直接进入XDB-C_(18)色谱柱分离并采用一级全扫描和数据依赖二级扫描(Full MS/dd MS~2)模式进行质谱分析.通过测定7-羟基香豆素的生成量评估5-甲氧基补骨脂素(5-MOP)对小鼠CYP2A5酶活性的影响.结果表明:各目标物的检出限为0.011~0.016 ng/m L,回收率和日内精密度分别为90.2%~110.4%和1.86%~7.08%.方法选择性好,分析效率高,适合于小鼠血液中香豆素及其代谢物的测定.5-MOP组小鼠香豆素的最大血药浓度Cmax为对照组的1.42倍(P0.01),药时曲线下面积AUC0-∞为1.59倍(P0.05),半衰期t_(1/2)为1.05倍(P0.01),5-MOP能够显著抑制小鼠CYP2A5酶活性.
[Abstract]:Using coumarin as the probe substrate, an ultra-high performance liquid chromatography-orbital trap high resolution mass spectrometry (UPLC-Orbitrap HRMS) method was established for the determination of coumarin and its metabolites in blood of mice. Using 4-methylpara-ketone as internal standard, the protein was precipitated with acetonitrile. It was separated directly into XDB-Cn18) column and analyzed by mass spectrometry using full MS/dd MS2) mode. The activity of 5-methoxy psoralen 5-MOP was evaluated by determining the production of 7-hydroxycoumarin in mice, and the activity of 5-methoxy-psoralen 5-MOP in mice was evaluated by measuring the content of 7-hydroxy-coumarin (7-hydroxy-coumarin). The results showed that the detection limit of each target was 0.011 ~ 0.016 ng/m / L, the recovery rate and intra-day precision were 90.22 ~ 110.4% and 1.86 ~ 7.08, respectively. Analysis efficiency is high, The maximum blood concentration of coumarin in 5-MOP group was 1.42 times of that of control group (P 0.01), the area under the curve was 1.59 times (P 0.05), and the half-life of coumarin was 1.05 times (P 0.01 / 2), and the activity of CYP2A5 was significantly inhibited by 5-MOP.
【作者单位】：中国烟草总公司郑州烟草研究院;
【基金】：国家自然科学基金(21175153,21307163) 中国烟草总公司郑州烟草研究院院长科技发展基金项目(602011CA0310)
【分类号】：R-33;O657.63

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