HPLC-MS法同时测定大鼠血浆中大环内酯类成分的浓度

Simultaneous determination of macrolide antibiotics in rat plasma by high performance liquid chromatography tandem mass spectrometry

目的:建立高效液相色谱串联质谱法测定大鼠血浆中红霉素、螺旋霉素Ⅰ、脱水红霉素、红霉素A烯醇醚、新螺旋霉素Ⅰ的浓度。方法:用乙腈提取,SCX固相萃取柱净化,上清液氮气吹干后甲醇复溶待测;5个大环内酯类成分通过Hypersil Gold C_(18)色谱柱(100 mm×2.1 mm,5μm)分离,质谱电喷雾正离子模式检测,多反应离子监测扫描,标准曲线法定量。结果:5个大环内酯类成分浓度在5~500 ng·mL^(-1)范围内的线性关系良好(r>0.999),在3个浓度水平下,5个大环内酯类成分的回收率可达84.6%~96.2%,精密度为1.6%~2.5%;5个大环内酯类成分的基质效应在81.23%~109.3%之间;方法检测下限为1.4~1.9μg·kg^(-1),方法定量下限为4.5~6.8μg·kg^(-1)。结论:HPLC-MS法测定大鼠血浆中5个大环内酯类成分的分析方法简单、快速,特异性强,适用于血浆中大环内酯类成分的测定。

Objective：To determine of erythromycin A,spiramycinⅠ,anhydroerythromycin A,erythromycin A enol ether,neospiramycinⅠin rat plasma by liquid chromatography-tandem mass spectrometry（HPLC-MS）method.Method：The macrolide components in samples were extracted with acetonitrile and purified by SPE with SCX culumn.The supernatant was evaporated to dryness under a stream of nitrogen and the residue obtained was redissolved in methanol.5 macrolide components were separated by Hypersil Gold C18（100 mm×2.1 mm,5μm）chromatographic column.Electro spray ionization was used to analyze these macrolide components by using positive ion mode.Multiple reaction monitoring in mass spectrometry was used to monitor ions.Standard calibration curve was used to quantitative calculation.Results：5 macrolide components showed good linear relationship in the range of 5-500 ng·mL^（-1）.The linear correlation coefficient were all greater than 0.999.At three concentration levels,the average recoveries were 84.6%-96.2%and the precisions were 1.6%-2.5%.The matrix effect of the 5 macrolides were between 81.23% and 109.3%.The limits of detection of 5macrolide components were 1.4-1.9μg·kg^（-1）,and the limits of quantitative were 4.5-6.8μg·kg^（-1）.Conclusion：The analytical method of 5 macrolide components in plasma by HPLC-MS is simple,rapid and specific.The analytical method is suitable for the determination of macrolide components in plasma.