LC-MS/MS法测定健康人体血浆中阿齐沙坦的浓度及在药代动力学研究中的初探

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t_max=2.22h，平均半衰期t_1/2=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) were monitored. 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:t_max=2.22 h, t_1/2=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.

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