UPLC-MS/MS测定大鼠舌下静脉注射阿西替尼的血药浓度及其药动学研究

Study on Plasma Concentration and Pharmacokinetics of Axitinib After Intravenous Administration in Rats by UPLC-MS/MS

关键词:  阿西替尼  超高效液相色谱串联质谱法  静脉给药

Key words:  Axitinib  UPLC-MS/MS  intravenous administration

摘要:

目的 建立UPLC-MS/MS对舌下静脉注射大鼠血浆中阿西替血药浓度的检测方法，并进行其药动学研究。方法 乙腈沉淀法处理样品，以伊马替尼为内标，Waters Acquity BEH C₁₈色谱柱（2.1 mm×100 mm，1.7 μm），流动相为0.1%甲酸水和乙腈。流速为0.3 mL·min^-1，柱温35℃。质谱条件为电喷雾电离源（ESI），检测方式为正离子电离、多离子反应监测（MRM）检测。结果 阿西替尼血药浓度在0.1~100 ng·mL^-1内线性关系良好（r=0.999 6），定量下限为0.1 ng·mL^-1。日内、日间精密度RSD<5.56%，提取回收率>87.87%。大鼠血浆中的阿西替尼在室温、4℃，-80℃，12 h后以及反复冻融3次后均有良好的稳定性，且不存在基质效应。结论 建立的测定阿西替尼血药浓度的方法准确、简单、快速，可以为阿西替尼的临床研究提供实验参考和依据。

Abstract:

OBJECTIVE To develop a ultra high-performance liquid chromatography-tandem mass spectrometry separation method(UPLC-MS/MS) for the pharmacokinetic study of Axitinib after intravenous administration. METHODS Plasma samples were treated by acetonitrile precipitation. The effective UPLC-MS/MS separation of the examined compounds was applied on an Acquity BEH C₁₈ column (2.1 mm×50 mm, 1.7 μm) column with a gradient mobile phase system consisting of 0.1% formic acid in water and acetonitrile. The analysis was performed with a flow rate of 0.3 mL·min^-1. An electrospray ionization (ESI) was used to detect in a positive ion mode. The scanning mode was MRM. RESULTS The assay was validated over concentration ranges of 0.1-100 ng·mL^-1, with a lower limit of quantification (LLOQ) of 0.1 ng·mL^-1. Intra-and inter-assay precision values for replicate quality control samples were within 5.56%. Assay recoveries of Axitinib was higher than 87.87%. Axitinib were stable in rat plasma for at least 24 h at room temperature, 30 days at 4℃ and -20℃, and following at least three freeze-thaw cycles. Furthermore, no notable matrix effect was observed for Axitinib. CONCLUSION The accurate and simple method developed can be provided the basis and applied to clinical pharmacokinetic study.