UPLC-MS/MS检测大鼠体内达卢生坦的血药浓度及其药动学研究

Study on Plasma Concentration and Pharmacokinetics of Darusentan in Rat Plasma by UPLC-MS/MS

关键词:  达卢生坦  UPLC-MS/MS  大鼠血浆  药动学

Key words:  darusentan  UPLC-MS/MS  rat plasma  pharmacokinetic

摘要:

目的 建立一种快速、高灵敏度和高选择性的UPLC-MS/MS用于大鼠血浆中达卢生坦的测定。方法 通过蛋白沉淀法完成血浆样品处理。通过Acquity UPLC BEH C₁₈柱（2.1 mm×50 mm，1.7 μm）色谱柱，流动相为0.1%甲酸水-乙腈，流速为0.4 mL·min^-1。采用ESI⁺方式检测，扫描方式为多反应离子监测方式检测。结果 达卢生坦血药浓度在0.01~2.5 μg·mL^-1内线性关系良好（R²>0.999 6），定量下限（LLOQ）为0.01 μg·mL^-1。血浆中的达卢生坦平均回收率为87.613%~97.115%，日内及日间精密度RSD<7.71%，基质效应RSD<9.34%。结论 该方法灵敏高，快速高效，能成功应用于大鼠口服10.0 mg·kg^-1达卢生坦后的药动学研究。

Abstract:

OBJECTIVE To develop a rapid, highly sensitive and selective UPLC-MS/MS for the determination of darusentan in rat plasma. METHODS Plasma samples were processed by protein precipitation. The separation was achieved by a gradient mobile phase, consisting of an aqueous solution of 0.1% formic acid and acetonitrile using an Acquity UPLC BEH C₁₈ column (2.1 mm×50 mm, 1.7 μm). The flow rate was 0.4 mL·min^-1. ESI⁺ was used to detect. The scanning mode was MRM. RESULTS The standard curve was linear over the range of 0.01-2.5 μg·mL^-1(R²>0.999 6) with a LLOQ of 0.01 μg·mL^-1. The average recovery of darusentan in plasma was 87.613%-97.115%. The RSD of intra-and inter-assay precision andaccuracy were <7.71%. The RSD of matrix effect was <9.34%. CONCLUSION This method is successfully applied in pharmacokinetic study after oral administration of 10.0 mg·kg^-1 darusentan in rats.