在线固相萃取-液相色谱串联质谱法测定阿姆西汀异构体的药代动力学差异  

Determination of Pharmacokinetics Differences of Ammuxetine Isomers in Rat Plasma Using On-Line Solid Phase Extraction with Liquid Chrommatography-Tandem Mass Spectrometry

作　　者：李迎[1] 冯航[1] 龚伟[1] 喻芳邻[1] 谢向阳[1] 何新华[1] 张有志[1] 梅兴国[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850

出　　处：《分析化学》2014年第12期1717-1722,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金青年基金资助项目(No.81102498)~~

摘　　要：建立了在线固相萃取-高效液相色谱-串联质谱法（ On-line SPE HPLC-MS/MS）测定大鼠血浆中S/R-阿姆西汀手性异构体的方法。血浆样品经过以下预处理步骤：采用甲醇-乙腈（50：50, V/V）沉淀蛋白；应用On-line SPE将血浆样品中的其它杂质去除；将保留在SPE柱上的阿姆西汀和内标洗脱后用分析柱进一步分离。分离后再用串联质谱法分别测定大鼠血浆中S/R-阿姆西汀的含量。 SPE柱为 Retain PEP Javelin（10 mm ×2.1 mm）；分析柱为ZORBAX SB-C18（50 mm ×2.1 mm×3.5μm）。质谱采用电喷雾离子源（ESI）,多反应监测（MRM）模式,检测离子为正离子,分别选择m/z 292.1/154.0和260.4/116.2作为S/R-阿姆西汀和内标（普萘洛尔）的检测离子对。结果表明,S/R-阿姆西汀的线性范围为0.2～1000μg/L,相关系数 R 分别为0.9903和0.9951,批内精密度 RSD 分别为1.2％～12.0％和0.4％～11.2％,回收率分别为94.2％～101.6％和94.3％～109.4％之间。本方法显著提高了阿姆西汀的检测灵敏度,可以满足大鼠灌胃给予阿姆西汀两种异构体的药代动力学研究要求。An on-line solid phase extraction （SPE）coupled with HPLC-MS/MS method was developed to determine S-ammuxetine and R-ammuxetine in rat plasma. The sample preparation consisted of the following steps： A protein precipitation extraction used methanol and acetonitrile （50：50, V/V）; an on-line SPE treatment to remove most matrixes in plasma; an enrichment and separation step used a Cls analytical column. S- and R-ammuxetine were determined by tandem mass spectrometry. The SPE column was a Retain PEP Javelin （10 mm×2.1 mm × 5μm） , while the chromatographic separation was achieved using a ZORBAX SB-C18 （50 mm x 2. 1 mmx 3.5 ~zm） analytical column with an isocratic mobile phase composed of acetonitrile-water-formic acid （40： 60： 0.1, V/V/V, 0.3 mL/min）. The selected reaction monitoring mode of the positive ion was performed and the precursor to the product ion transitions of m/z 292. 1/154.0 and m/z 260.4/116.2 were used to measure S-ammuxetine, R-ammuxetine and internal standard （propranolol）. The method was linear over a concentration range from 0.2 to 1000 μg/L with the correlation coefficients of 0.9903 and 0. 9951. The average intra-day precision values were 1.2% -12.0% for S-ammuxetine and 0.4%-11.2% for R-ammuxetine, respectively. The average recoveries were 94.2%-101.6% for S- ammuxetine and 94. 3%- 109.4% for R-ammuxetine. Compared to the literature, the sensitivity of this method increased dramatically. The present method has been successfully applied to the preclinical rat research of ammuxetine isomers following intragastric administration.

关 键 词：在线固相萃取-液相色谱质谱联用 阿姆西汀手性异构体 大鼠药代动力学 新型抗抑郁药 

分 类 号：R965[医药卫生—药理学] O657.63[医药卫生—药学]