高通量甲基衍生化固相萃取LC-MS/MS法定量测定人血浆内的米诺膦酸  

High-throughput On-cartridge Methyl-derivatization Coupled with Solid-phase Extraction for Quantitative Determination of Minodronic Acid in Human Plasma by LC-MS/MS Method

作　　者：韩茉 邹巧根[1] 陈茜睿 潘诗苑 HAN Mo;ZOU Qiaogen;CHEN Xirui;PAN Shiyuan(College of Biological and Pharmaceutical Engineering,Nanjing Technology University,Nanjing 211800)

机构地区：[1]南京工业大学生物与制药工程学院

出　　处：《中国医药工业杂志》2020年第1期102-107,共6页Chinese Journal of Pharmaceuticals

摘　　要：采用氘代米诺膦酸为内标,建立了高通量衍生化固相萃取LC-MS/MS的定量方法检测人血浆中的米诺膦酸(1)。将血浆样品300μl上样于弱阴离子交换96孔固相萃取板上,配合正压装置在pH 7.0避光条件下与三甲基硅烷化重氮甲烷(TMS-DAM)发生衍生化反应,洗脱液于烘箱70℃反应1 h使衍生化完全。以Eclipse XDB苯基柱为色谱柱,流动相为乙腈∶含0.15%甲酸10 mmol/L乙酸铵溶液,线性梯度洗脱。采用电喷雾离子源(ESI),正离子模式,以多反应监测(MRM)扫描方式进行监测,用于定量分析的离子对为m/z 393.2→m/z 267.1(五甲基米诺膦酸)和m/z 397.1→m/z271.2(D4-五甲基米诺膦酸)。本法线性范围为10~1 500 pg/ml,批内RSD为2.6%~7.5%,批间RSD为2.5%~7.9%;本法提取回收率为89.7%~96.8%,基质效应为96.9%~102.3%。本方法灵敏、快速、高通量,可用于人血浆中1的含量测定和临床研究。A quantitative method for determination of minodronic acid(1) in human plasma was established by using D4-minodronic acid(2) as the internal standard. Plasma sample(300 μl each) was deposited on the weak anion-exchange 96-well solid phase extraction plate, and the eluent was completely reacted with the trimethylsilanized diazomethane(TMS-DAM) at 70 ℃ for 1 h away from bright light at pH 7.0 in a positive pressure device. The Eclipse XDB phenyl column was used with the mobile phase of acetonitrile∶10 mmol/L ammonium acetate aqueous solution containing 0.15% formic acid in a linear gradient elution mode. Electrospray ionization source(ESI) and positive ion mode was selected. Multi-reaction ion monitoring(MRM) scanning was used for monitoring. The ion pairs set for quantitation were m/z 393.2→m/z 267.1(pentamethylated 1) and m/z 397.1→m/z 271.2(pentamethylated 2), respectively. The calibration curve of 1 showed good linearity over the range of 10-1 500 pg/ml. The inter-batch RSD was 2.6%-7.5%, and the intra-batch RSD was 2.5%-7.9%. The extraction recovery was 89.7%-96.8%, and the matrix effect was 96.9%-102.3%. This method was successfully used to characterize the pharmacokinetic profile of 1 in healthy volunteers(n=12), indicating it could be used for the determination of 1 in human plasma and clinical research.

关 键 词：米诺膦酸 衍生化 固相萃取 液质联用 体内药物分析 

分 类 号：O657.63[理学—分析化学]