液相色谱-串联质谱法快速测定人血浆中乐福昔布的含量

Determination of Lefucoxib in Human Plasma by Liquid Chromatography-tandem Mass Spectrometry

建立测定人血浆中乐福昔布含量的LC-MS/MS方法。取正常人血浆250μL,然后加入200ng/mL内标塞来昔布50μL,加入1mL无水乙醚涡旋震荡提取2min,离心10分钟(12000转/分)后,取上清液,下层再加入乙醚,同法再萃取一次,合并2次萃取液,吹干溶剂后,用30%的乙腈250μL溶解,取10μL进行LC-MS/MS测定。LC条件:采用ASB C18柱(2.1×50mm,5μm),流动相:乙腈-0.1%甲酸梯度洗脱,流速:0.4mL/min。质谱条件:ESI电离源,正离子模式,多反应监测(MRM)方式,用于定量分析的离子对分别为m/z 395.1→m/z 296.1(乐福昔布)和m/z 382.2→m/z 281.0(内标,塞来昔布)。该方法乐福昔布的线性范围为0.4～800 ng/mL,最低检测限为0.2ng/mL。本方法操作简便、快速、结果准确、可用于该药物的含量测定,同时也可为临床药代动力学研究提供参考。

A LC-MS/MS method to fast determine lefucoxib in human plasma was established.The internal standard（celecoxib） 50μL was added to plasma,then it was extracted by 1mL diethyl ether for two minutes.After centrifugelling and transfering and evaporating,the residue was dissolved with 250μL 30% acetonitrile and was determined by injecting 10μL to LC-MS/MS system.ASB C18column was used and the mobile phase consisted of acetonitrile--0.1% formic acid gradient system at the flow rate of 0.4mL/min.MS determination was in electrospray ionization（ESI） positive mode and in multi-reaction monitoring（MRM） mode.The ion pair in quantitative analysis was m/z 395.1→m/z 296.1（lefucoxib）,m/z 382.2→m/z281.0（celecoxib,internal standard）,respectively.The linear range was 0.4～800 ng/mL（r =0.9974）,and the limit of detection for lefucoxib was 0.2 ng/mL.The method is simple,rapid,sensitive,and reliable,and it is suitable for the determination of drug concentration of lefucoxib in human plasma.It also can provide some reference for clinical pharmacokinetics research.