LC-MS/MS法测定Beagle犬血浆中ET-26盐酸盐及其代谢产物ET-acid

Determination of ET-26 hydrochloride and its metabolite ET-acid in plasma of Beagle dogs by LC-MS/MS method

该文通过建立LC-MS/MS方法,采用全扫描离子(MRM)检测方式,测定静脉注射给药后Beagle犬中ET-26盐酸盐及其代谢产物依托咪酯酸的含量。采用色谱柱Agilent Extend-C18(3.0 mm×100 mm,3.5μm),流动相为水(0.01%NH3,用乙酸调pH至9.0)-乙腈,体积比35:65,采用ESI离子源,正离子模式下定量分析碎片离子质荷比(m/z)有ET-26盐酸盐275.3/105.1,依托咪酯酸m/z 217.0/113.1,依托咪酯杂质C(内标)m/z 259.0/155.1。ET-26盐酸盐与依托咪酯酸分别在3.225~2 090 ng/mL、3.253~2 108 ng/mL范围内,线性关系良好;定量限分别为3.225 ng/mL、3.253 ng/mL;检测限分别为0.97 ng/mL、1.00 ng/mL;日内、日间精密度RSD均小于15%;回收率分别在86.57%~105.03%、95.94%~98.27%范围内。该方法成功应用于Beagle犬中ET-26盐酸盐的药代动力学参数测定。结果表明该方法可以灵敏、有效地测定Beagle犬血浆中ET-26盐酸盐及其代谢产物依托咪酯酸的含量。

In this study, a method was established to determine the plasma concentration of ET-26 hydrochloride and and its metabolite etomidate acid（ET acid） in Beagle dogs by using LC-MS/MS. Agilent Extend-C18（3.0 mm×100 mm, 3.5 μm） was used as the column. The mobile phase A was 0.01% ammonium solution adjused to pH 9 with acetic acid, and the mobile phase B was acetonitrile with volume ratio of 35：65.The detection was performed by electrospray ionization（ESI） mass spectrometry using multi-reactions monitoring mode（MRM） in positive ion mode while the precursor-to-product ion pairs were at m/z 275.3/105.1 for ET-26 hydrochloride, m/z 217.0/113.1 for etomidate acid and m/z 259.0/155.1 for the IS. Good linearity was generated over the concentration range of 3.225-2 090 ng/mL with the limit of detection and quantificationof 0.97 ng/mL and 3.225 ng/mL, respectively for ET-26 hydrochloride. While for etomidate acid, good linearity was generated over the concentration range of 3.253-2 108 ng/mL with the limit of detection and quantification of 1.00 ng/mL and 3.253 ng/mL. Intra-day and inter-day precisions RSD were less than 15% and the mean recoveries ranged from 86.57%-105.03%, 95.94%-98.27%, respectively. The developed method was successfully applied to the determination of pharmacokinetic parameters of ET-26 hydrochloride in Beagle dogs. The developed method was sensitive, suitable for determining the ET-26 hydrochloride and its metabolite ET-acid in plasma of Beagle dogs.