UPLC-MS/MS检测大鼠血浆中阿帕替尼的浓度及其药动学研究

Concentration Determination of Apatininb in Rat Plasma by UPLC-MS/MS and Its Application to Pharmacokinetics Study

关键词:  阿帕替尼  超高效液相色谱串联质谱法  浓度  药动学

Key words:  apatinib  UPLC-MS/MS  concentration  pharmacokinetics

摘要:

目的 采用UPLC-MS/MS建立快速检测大鼠血浆中阿帕替尼浓度的方法，并应用于药动学研究。方法 大鼠血浆样本用乙腈沉淀蛋白，液质联用技术检测浓度，流动相为乙腈-水（含0.1%甲酸），梯度洗脱，流速为0.3 mL·min^-1，柱温40℃，内标为氯唑沙宗；质谱条件：电喷雾离子化源（ESI），负离子监测模式，检测离子对阿帕替尼为m/z 396.2→210.0和m/z 396.2→158.0，氯唑沙宗m/z 168.0→132.0。结果 阿帕替尼和内标氯唑沙宗的保留时间分别为1.07 min和1.40 min，线性范围为10~2 000 ng·mL^-1（r²=0.993），检测限为1 ng·mL^-1，准确度为90.65%~111.50%，基质效应为89.14%~104.65%，平均回收率>86%，日内、日间精密度RSD均<10%。常温下放置24 h、冻融2次和-80℃冻存30 d的RSD均<10%。药动学研究结果显示，大鼠单次灌胃阿帕替尼76.5 mg·kg^-1，AUC_（0-t）为（6 114.41±645.99）ng·mL^-1·h，CL_z/F为（12.21±1.08）L·h^-1·kg^-1，V_z/F为（75.70±38）L·kg^-1，T_1/2为（4.23±1.94）h，T_max为（2±0.71）h，C_max为（1 377.7±284.54）μg·L^-1。结论 该法操作简便，重复性好，准确可靠，适用于大鼠血浆中阿帕替尼的浓度检测及其药动学研究。

Abstract:

OBJECTIVE To establish a UPLC-MS/MS method for rapid determination of apatinib concentration in rat plasma and apply to pharmacokinetics study. METHODS Rat plasma samples were prepared with acetonitrile to precipitate the protein. The apatinib concentration in rat plasma was determined by liquid chromatography-mass spectrometry. The mobile phase was acetonitrile-water (containing 0.1% formic acid). Gradient elution with a flow rate of 0.3 mL·min^-1, column temperature was 40℃, chlorzoxazone was used as internal standard. Mass spectrometry conditions:Electrospray ionization source(ESI), negative ion detection mode, detection pairs were m/z 396.2→210.0 and m/z 396.2→158.0 for apatinib, m/z 168.0→132.0 for chlorzoxazone. RESULTS The retention time of apatinib and internal standard chlorzoxazone were 1.07 min and 1.40 min, respectively. The linear range was 10 to 2 000 ng·mL^-1 (r²=0.993) and the limit of quantification was 1 ng·mL^-1. The accuracy of the method was in the range of 90.65%-111.50%, and the matrix effect was 89.14%-104.65%. Mean recoveries of apatinib in rat plasma were >86%. RSD of intra-day and inter-day precision were both <10%. The RSDs of stabilities of 24 h kept in room temperature, froze-thaw 2 times, 30 d froze in -80℃ were all<10%. Pharmacokinetic study showed that after the rats received a single administration of apatinib with 76.5 mg·kg^-1, the AUC_(0-t) was (6 114.41±645.99) ng·mL^-1·h, and CL_z/F was (12.21±1.08) L·h^-1·kg^-1, V_z/F was (75.70±38) L·kg^-1, T_1/2 was (4.23±1.94) h, T_max was (2±0.71) h, C_max was (1 377.7±284.54)μg·L^-1. CONCLUSION The method established is easy to operate, reproducible, accurate and reliable. It is suitable for the determination of apatinib concentration in plasma and can applied to its pharmacokinetics study.