UPLC-Q Exactive轨道阱高分辨质谱法测定小鼠血液中的NNN及其7种代谢物

为了准确测定小鼠血液中的N-亚硝基降烟碱(NNN)及其代谢物,通过对色谱、质谱条件的优化,建立了以NNN-d₄和HPB-d₄为内标,用乙腈沉淀蛋白后,直接经超高效液相色谱(UPLC)仪器的亲水相互作用色谱(HILIC)柱进行分离,通过全扫描/数据依赖二级质谱扫描(FS/dd MS2)测定小鼠血浆中NNN及其7种代谢物的UPLC-Q Exactive轨道阱高分辨质谱法。结果表明:①NNN及其7种代谢物的检出限为0.019~0.067 ng/m L,回收率在78.0%~117.5%之间,日内精密度为0.47%~8.38%。②该方法适合于小鼠血浆中NNN及其7种代谢物的同时检测。 

Determination of NNN and its Seven Metabolites in Mouse Blood with UPLC-Q Exactive Orbit Trap High-Resolution Mass Spectrometry

In order to accurately determine the N-nitrosonornicotine(NNN) and its metabolites in mouse blood,a UPLC-Q Exactive orbit trap high-resolution mass spectrometry method was established by optimizing chromatographic and mass spectrometric conditions and using NNN-d₄ and HPB-d₄ as internal standards.Wherein,after protein precipitating with acetonitrile,the sample was separated directly by an HILIC column in UPLC spectrometer.NNN and its metabolites in mouse plasma were analyzed by mass spectrometry under full scan/data-dependent MS2(FS/ddMS2) mode.The results showed that:1) The limits of detection and the recoveries of NNN and its metabolites ranged from 0.019 to 0.067 ng/mL and from 78.0% to 117.5%,respectively,with the intra-day precision of 0.47%-8.38%.2) The developed method was suitable for the simultaneous analysis of NNN and its seven metabolites in mouse plasma.