UPLC-MS/MS测定大鼠血浆中奥拉帕利的浓度及其药动学研究

Determination of Olaparib in Rat Plasma by UPLC-MS/MS and Its Pharmacokinetic Study

目的建立一种具有高灵敏度和高选择性测定大鼠血浆中奥拉帕利药物浓度的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法,并研究奥拉帕利在大鼠体内的药动学特征。方法大鼠血浆样品采用乙腈沉淀法去除蛋白,色谱柱为Acquity UPLC BEH C_(18)柱(2.1 mm×50 mm,1.7μm),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱。质谱采用电喷雾离子源,多反应监测正离子模式,奥拉帕利定量离子对为m/z 435.19→m/z 367.1。以50 mg·kg^(-1)奥拉帕利经大鼠灌胃给药后于不同时间点采集血样,利用建立的方法进行检测分析,并用DAS2.0软件计算药动学参数。结果血浆中奥拉帕利在(0.6~1821)ng·mL^(-1)内线性关系良好,定量限为0.15 ng·mL^(-1),日内、日间精密度RSD均小于5.5%,准确度在95.4%~99.2%内,平均提取回收率为84.2%~96.0%,基质效应在91.4%~108.8%内。结论建立的方法灵敏度高、结果准确,可用于大鼠血浆中奥拉帕利质量浓度的测定及其药动学研究。

OBJECTIVE To establish a ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method with high sensitivity and high selectivity for determination of olaparib in rat plasma,and study the pharmacokinetics of olaparib.METHODS The protein of rat plasma was removed by acetonitrile precipitation method.The analytical column was Acquity UPLC BEH C_(18) column(2.1 mm×50 mm,1.7μm).The mobile phase consisted of 0.1%formic acid and acetonitrile by gradient elution.The electrospray ion source with positive ion detection was used for the detection.Multiple reaction monitoring was used to monitor the precursor-to-product ion transitions of m/z 435.19→m/z 367.1 for olaparib.The blood samples were collected in the different time after intragastric administration to rats at 50 mg·kg^(-1),and measured by established UPLC-MS/MS method.The pharmacokinetic parameters were calculated by DAS 2.0.RESULTS The linear range of olaparib in plasma was(0.6-1821)ng·mL^(-1),the lower limit of quantification was 0.15 ng·mL^(-1),intra-day and inter-day precisions were less than 5.5%,the accuracy were between 95.4%and 99.2%,the average extract recoveries ranged from 84.2%to 96.0%,and the matrix effect were between 91.4%and 108.8%.CONCLUSION The established UPLC-MS/MS method is sensitive,accurate,and it has been successfully applied to the determination of olaparib in rat plasma and its pharmacokinetic study.