LC-MS/MS测定阿齐沙坦及其盐在大鼠血浆中的药动学研究

Study on Pharmacokinetics of Azilsartan and Its Salt in Rat Plasma by LC-MS/MS

关键词:  阿齐沙坦  盐  大鼠血浆  液相色谱-串联质谱法  药动学

Key words:  azilsartan  salt  rat plasma  LC-MS/MS  pharmacokinetics

摘要:

目的 建立液相色谱-串联质谱法测定大鼠血浆中阿齐沙坦及其盐的浓度并研究其药动学。方法 大鼠血浆样本以乙腈沉淀蛋白后，采用Eclipse Plus C₁₈色谱柱（50 mm×3.0 mm，1.8 μm）；流动相（乙腈：水=60：40），流速为0.35 mL·min^-1，柱温为40℃；采用Agilent 6430三重四极杆串联质谱仪，离子化方式：电喷雾-正离子（API-ES）；监测方式：MRM；阿齐沙坦监测离子对457.3/233.1，缬沙坦监测离子对436.2/291.4，用作内标。SD大鼠灌胃给予阿齐沙坦1.0 mg·kg^-1及阿齐沙坦盐1.2 mg·kg^-1。结果 阿齐沙坦在5~30 000 ng·mL^-1内线性关系良好；回收率为85%~115%，精密度RSD在±15%内。阿齐沙坦盐大鼠体内主要动力学参数如下：AUC_{（0-24 h）}为（12.9±3.2）μg·mL^-1·h^-1，AUC_（0-∞）为（14.2±4.1）μg·mL^-1·h^-1，C_max为（3.8±0.3）μg·mL^-1，T_1/2为（13.4±0.5）h。阿齐沙坦的主要动力学参数如下：AUC_{（0-24 h）}为（8.1±2.6）μg·mL^-1·h^-1，AUC_（0-∞）为（9.7±3.1）μg·mL^-1·h^-1，C_max为（2.3±0.5）μg·mL^-1，T_1/2为（10.5±0.5）h。结论 本法经方法学验证，适用于大鼠血浆中阿齐沙坦及其盐的浓度测定，可用于阿齐沙坦及其盐大鼠体内药动学研究。

Abstract:

OBJECTIVE To establish an LC-MS/MS method to determine azilsartan and its salt in rat plasma and investigate their pharmacokinetics. METHODS After protein precipitation with acetonitrile, the analytes and internal standard were separated on Eclipse Plus C₁₈ column(50 mm×3.0 mm, 1.8 μm) with acetonitrile-water (60:40) as mobile phase eluted at a flow rate of 0.35 mL·min^-1. Detection was carried out by electrospray positive ionization mass spectrometry in the multiple reaction monitoring(MRM) mode. The MRM transitions of m/z 457.3/233.1 and m/z 436.2/291.4 were used to quantify azilsartan and valsartan, respectively. SD rats were given azilsartan of 1.0 mg·kg^-1 while the salt of 1.2 mg·kg^-1. RESULTS The calibration curve of azilsartan was linear over the concentration range of 5-30 000 ng·mL^-1. The RSDs of accuracy were <15%. The main pharmacokinetics parameters of the salt in rats were estimated as follows:AUC_{(0-24 h)}was (12.9±3.2)μg·mL^-1·h^-1, AUC_(0-∞) was (14.2±4.1)μg·mL^-1·h^-1, C_max was (3.8±0.3)μg·mL^-1, T_1/2 was (13.4±0.5)h. The main pharmacokinetics parameters of azilsartan were estimated as follows:AUC_{(0-24 h)} was (8.1±2.6)μg·mL^-1·h^-1, AUC_(0-∞) was (9.7±3.1)μg·mL^-1·h^-1, C_max was (2.3±0.5)μg·mL^-1, T_1/2 was (10.5±0.5)h. CONCLUSION The established method can be applied on the determination of azilsartan and its salt in plasma of rats and is suitable for the pharmacokinetics study.