LC-MS/MS法测定大鼠血浆中利巴韦林及其毒代动力学参数

Determination of Ribavirin in Rat Plasma and Its Toxicokinetic Parameters by LC-MS/MS

目的:建立LC-MS/MS法,测定大鼠血浆中利巴韦林及其毒代动力学参数。方法:LC条件:Waters Atlan-tis T3色谱柱(4.6 mm×150 mm,3μm),柱温为40℃,流动相为0.1%甲酸水溶液-0.1%甲酸乙腈溶液(95∶5),流速为1 000μL.min-1,进样量为2μL;MS条件:电喷雾离子源,正离子电离模式,多反应离子监测的MS扫描方式。以10、40、160 mg.kg-1.d-1剂量每日1次、连续28 d给大鼠灌胃使用利巴韦林药液,测定其血药浓度和毒代动力学参数。结果:利巴韦林血药浓度线性范围为5～5 000μg.L-1,最低定量限为5μg.L-1;日内和日间精密度良好,RSD均小于15%,准确度均在±15%以内。所测利巴韦林毒代动力学参数基本上与给药剂量成正相关,未见有明显药物蓄积现象,与毒性表现基本一致。结论:该法专属性强、灵敏度高、操作简便,适用于生物样本中利巴韦林的测定及毒代动力学研究。

Objective： To establish a LC-MS/MS method for the quantitative determination of ribavirin in rat plasma and its toxicokinetic parameters. Methods： A Waters Atlantis T3 column （4. 6 mm x150 mm, 3 txm） at a temperature of 40 ℃ and 0. 1% formic acid in water - 0. 1% formic acid in acetonitrile （95： 5） as mobile phase at a flow rate of 1 000 μL-min-1 were used with a injection volume of 2 μL for LC. ESI and MRM were used with positive ionization model for MS. Ribavirin （10, 40 and 160 mg-kg-1, po, qd, for 28 d） was administered in rats and its plasma concentration and toxicokinetic profile were determined. Results： The calibration curve for ribavirin had a good linearity over the range of 5 - 5 000μg-L-1 and the minimum quantitative limit was 5 μg.L-1. The precisions （RSDs） of inter- and intra-day and the accuracies （REs） were below 15% and ＋ 15% respectively. The measured toxicokinetic parameters of ribavirin were correlated positively with oral dose and indicated no significant drug accumulation. Conclusion The method is specific, sensitive, convenient and suitable for the determination of ribavirin in biological samples and its toxicokinetic study.