LC-MS/MS测定大鼠血浆中4-氯-2-氟-3-甲氧基苯硼酸及其毒代动力学研究

Determination of 4-Chloro-2-fluoro-3-methoxybenzeneboronic Acid in Rat Plasma by LC-MS/MS and Its Application in the Toxicokinetics Study

目的建立一种快速灵敏的高效液相色谱串联质谱法(LC-MS/MS)用于大鼠血浆中4-氯-2-氟-3-甲氧基苯硼酸(4-chloro-2-fluoro-3-methoxybenzeneboronic acid,PBA)的测定,并进行PBA的毒代动力学研究。方法观察SD大鼠分别灌胃及静脉注射染毒不同剂量PBA后血浆中PBA的浓度变化。采用内标法进行定量分析,获得血浆中PBA的浓度,并采用DAS药动学软件拟合主要动力学参数AUC、Cmax、Tmax、T1/2等。结果血浆中PBA提取回收率为88.8%~94.4%,日间精密度为6.4%~8.9%,日内精密度为3.9%~5.9%,准确度为97.8%~105.5%。PBA的线性范围为20~2 500 ng·m L-1,对照品曲线的相关系数r>0.99。PBA的定量下限为20 ng·m L-1(RSD=9.67%,n=5)。结论该方法能成功应用于PBA在大鼠体内的血浆毒代动力学。PBA在大鼠体内的暴露水平未随着染毒剂量的增加而线性增加,表明PBA在大鼠体内的吸收呈现饱和性。

OBJECTIVE To develop a rapid, highly sensitive and selective high performance liquid chromatography tandem mass spectrometry（LC-MS/MS） for the determination of 4-chloro-2-fluoro-3-methoxybenzeneboronic acid（PBA） in rat plasma. Explore and complete the toxicokinetic study of PBA in rats. METHODS To observe the plasma concentration change of PBA in SD rats after treated with different doses of PBA orally and intravenously, respectively. Internal standard method was used in analysis to quantify the PBA plasma concentration. DAS software was used to obtain kinetic parameters such as AUC, Cmax, Tmax, T1/2 and so on. RESULTS The extraction recovery rate of PBA in rat plasma was in the range of 88.8%–94.4%, the intra-day precision was in the range of 6.4%–8.9%, and the inter-day precision was in the range of 3.9%–5.9%, respectively. Accuracy was in the range of 97.8%–105.5%. Linear calibration curves for PBA was observed at 20–2 500 ng·m L-1 with correlation coefficient r0.99. The lowest limit of quantification（LLOQ） for PBA was 20 ng·m L-1（RSD=9.67%, n=5）. CONCLUSION This method is successfully applied in the rat plasma concentration determination of PBA. The plasma concentrations of PBA in rats has nonlinear dynamics and its systemic exposure did not raise as the administered doses.