摘要:
目的:为临床研究阿齐沙坦人体药动学建立一种高效、专一的液相色谱-串联质谱(LC-MS/MS)方法,并将其应用于阿齐沙坦在中国健康人群药动学研究.方法:含阿齐沙坦血浆样本与内标混合后经乙腈处理,以含5 mmol·L-1甲酸铵溶液-乙腈为流动相,SB-Aq色谱柱(3.0 mm×100 mm,3.5 μm)为分析柱,采用API 4000型液质联用系统,流速0.6 mL· min-1,室温下测定;采用负离子模式,MRM扫描,阿齐沙坦m/z 455.2→411.2和内标奥美拉唑m/z 344.1→193.9.9例健康受试者口服40 mg阿齐沙坦片,于不同时间点分别采集静脉血进行药代动力学分析.结果:阿齐沙坦和内标的保留时间分别为4.17和4.77 min,标准曲线在0.01~10.0 mg· mL-1范围内线性良好(r=0.998 6±0.000 9),最低定量限10 ng·mL-1,日内、日间批间差异均小于12%,准确度在89.2%~ 110.2%,回收率在83.2%~96.2%之间,所有的稳定性考察项目均符合要求.药动学结果显示,健康受试者口服阿齐沙坦片40 mg后,阿齐沙坦在人体内平均达峰时间tmax=2.22h,平均半衰期t1/2=10.15 h.结论:本测定方法适用于阿齐沙坦的药动学研究和血药浓度监测.%Objective:To develop a sensitive and specific method for the determination of azilsartan pharmacokinetics in clinical human study using high performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) system,and to apply for its pharmacokinetic feature in Chinese healthy volunteers.Methods:After mixed with internal standard (IS),the plasma samples were precipitated by acetonitrile,and the supernatant was detected with an API 4000 LC-MS/MS system,using SB-Aq (3.0 mm × 100 mm,3.5 μm)column with mobile phase 5 mmol· L-1 ammonium formate in water (A)-acetonitrile (B) at a flow rate of 0.6 mL· min-1 at room temperature.Negative electrospray ionization and multiple reation monitoring (MRM) mode were used.The transition of m/z 455.2 → 411.2 for azilsartan and m/z 422.1 → 198.0 for omeprazole (IS) weremonitored.Venous blood samples for pharmacokinetic measurements were collected at different time points from nine healthy volunteers with orally administering 40 mg of azilsartan.Results:The retention time of azilsartan and internal standard were 4.17 and 4.77 min,respectively.The standard curve was linear (0.01-10.0 μg· mL-1) with a good correlation coefficient (r=0.998 6 ± 0.000 9).The lower limit of quantification was 10 ng· mL-1.Intra-and inter-day assay variations were less than 12%,the accuracy values were between 89.2% and 110.2%,extraction recoveries ranged from 83.2% to 96.2% across the calibration curve range,and all stability tests met the acceptance criteria.Mean pharmacokinetic parameters for azilsartan in healthy volunteers after 40 mg of azilsartan were followed:tmax=2.22 h,t1/2=10.15 h.Conclusion:The established LC-MS/MS method is suitable for the pharmacokinetic study of azilsartan and the monitoring of blood drug level.