高效液相色谱-串联质谱法测定小鼠组织中的新型抗心力衰竭化合物AF-HF001

建立了基于高效液相色谱-质谱联用技术的新型抗心力衰竭化合物AF-HF001在小鼠组织中的测定方法,并考察了其在小鼠体内的组织分布。采用液-液萃取对样品进行预处理,以Thermo Hypersil Gold C18色谱柱进行分离,流动相为乙腈-0.1%甲酸水溶液,梯度洗脱,采用选择性反应监测(SRM)正离子模式检测分析物。小鼠组织中AF-HF001含量在0.5~10μg/mL范围内线性关系良好(r>0.996),在加标量为1.5,4,10μg/mL的组织样品中,提取回收率大于58%,基质效应在100.38%~109.99%之间,日内和日间精密度RSD均小于15%。该方法为AF-HF001的体内分布研究及进一步药物研发提供了依据。初步结果表明,该化合物能够迅速到达心脏并迅速代谢,主要分布在肝脏组织中,其他组织中分布较少。

Determination of AF-HF001 in mouse tissue by LC-MS/MS

A method based on high performance liquid chromatography-tandem mass spectrometry(LC-MS/MS)was developed for the determination of AF-HF001 in mice tissues,and the distribution of AF-HF001 in mice tissue was then investigated.The samples were pretreated by liquid-liquid extraction.The separation was performed on a Thermo Hypersil Gold C18(2.1 mm×50 mm,3μm)column with a gradient mobile phase consisting of acetonitrile and 0.1%aqueous formic acid.The analyte was detected by selective reaction monitoring(SRM)in positive ion mode.In mice tissues,the linear range of AF-HF001 was 0.5-10μg/mL(r>0.996).The extraction recoveries of AF-HF001 were more than 58%at spiked levels of 1.5(low),4(medium)and 10μg/mL(high)in mice tissues,respectively,and the matrix effect was between 100.38%and 109.99%.The intra-and inter-day precisions were less than 15%.This compound can reach the heart quickly,and distribute mainly in the liver tissue,and less in other tissues.In summary,the method provides effective reference information for the distribution and further drug development of AF-HF001.