UPLC-Orbitrap HRMS法测定小鼠血液中香豆素及其代谢物

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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-C18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 MS2)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0.0110.016 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(P<0.01),药时曲线下面积AUC0-∞为1.59倍(P<0.05),半衰期t1/2为1.05倍(P<0.01),5-MOP能够显著抑制小鼠CYP2A5酶活性.

In order to accurately determine coumarin and its metabolite in mouse blood, a UPLC-Orbitrap high resolution mass spectrometry method with coumarin as the probe substrate was established. Using 4-methylumbelliferone as internal standard, immediately after protein precipitating with acetonitrile, the samples were separated by an XDB-Ci8 column in UPLC spectrometer with mass spectrometry under full scan/data-dependent MS2 mode. The effect of 5-Methoxypsoralen （5-MOP） on the activity of cytochrome P450 2A5 （CYP2A5） enzyme in mouse was evaluated by the production of 7-hydroxycoumarin. The results showed that the limits of detection of analytes ranged from 0. Oil to 0. 016 ng/mL, and the recoveries ranged from 90. 2% to 110.4% with the intra-day precision of 1. 86% to 7. 08% respectively. The developed method was suitable for the simul-taneous analysis of coumarin and its metabolite in mouse blood with good selectivity and high efficiency. Compared with the control group, the maximum plasma concentration of 5-MOP-treated group was increased by 1. 42 fold （P〈C0. 01） , the area under the curve was increased by 1. 59 fold （P〈C0. 05） , the elimination half-lie was increased by 1. 05 fold （P〈C0. 01）. The re-sults indicate that the activity of CYP2A5 is markedly inhibited by 5-MOP.

作者:

李琛王荣浩王丁众毛健刘帅东张建勋

Li Chen Wang Ronghao Wang Dingzhong Mao Jian Liu Shuaidong Zhang Jianxun(Zhengzhou Tobacco Research Institute of CNTC, Zhengzhou 450001, China)

机构地区:

中国烟草总公司郑州烟草研究院

出处:

《betway官方app 学报：自然科学版》 CAS 北大核心 2017年第2期38-42,共5页

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)

关键词:

香豆素 5-MOP CYP2A5 小鼠血液 Orbitrap高分辨质谱

coumarin 5-methoxypsoralen CYP2A5 mouse blood Orbitrap high resolution mass spectrometry