液-质联用法测定大鼠血浆中异烟肼及其代谢产物乙酰异烟肼的浓度

Development of LC-MS/MS method for determination of isoniazid and acetylisoniazid in rat plasma

目的:建立应用LC-MS/MS测定大鼠血浆中异烟肼及其体内代谢物乙酰异烟肼浓度的方法,并将其应用于异烟肼及乙酰异烟肼在大鼠体内过程的研究。方法:血浆样品采用甲醇沉淀蛋白进行提取,以ESI电离源在正离子模式下检测,用Waters Cortecs C_(18)(4.6 mm×100 mm,2.7μm)色谱柱,甲醇-水溶液为流动相等度洗脱,流速为0.3 mL·min^(-1)。结果:异烟肼及乙酰异烟肼在大鼠血浆中的线性范围及定量限均符合测定要求,日内、日间精密度及基质效应均<15%。结论:本方法快速、灵敏、专属性强且重现性好,可用于大鼠血浆中异烟肼及乙酰异烟肼药动学研究。

OBJECTIVE To establish a LC-MS/MS method for determination of isoniazid and acetylisoniazid,its metabolite in vivo in rat plasma,and to apply the method to the study of pharmacokinetics of isoniazid and acetylisoniazid in rats.METHODS The plasma samples were extracted with protein precipitated with methanol and detected with ESI ionization source in positive mode.The samples were eluted with Waters Cortecs C18(4.6 mm×100 mm,2.7μm)column and methanol-water solution as mobile phase at a flow rate of 0.3 mL·min-1.RESULTS The linear range and quantitation limit of isoniazid and acetyl-isoniazid in rat plasma were consistent with the requirements of the assay,and the inter-,intra precision and matrix effect were<15%.CONCLUSION The method is rapid,sensitive,specific and reproducible for determination of isoniazid and aceylisoniazid in rat plasma.