LC-MS/MS法测定小鼠血浆nodosin浓度及其药动学研究  

Quantitative Analysis of Nodosin in Mice Plasma with LC-MS/MS Method and Pharmacokinetic Study of Nodosin

作　　者：任灵芝[1] 张丽媛[1] 王腾华[1] 董昕[1] 高幼衡[1] 

机构地区：[1]广州中医药大学中药学院,广东广州510006

出　　处：《中药新药与临床药理》2015年第2期205-209,共5页Traditional Chinese Drug Research and Clinical Pharmacology

摘　　要：目的建立LC-MS/MS法测定nodosin在小鼠体内的血药浓度,研究nodosin在小鼠体内的药物动力学。方法将KM小鼠单次腹腔注射nodosin生理盐水注射液,断头取血,采用液液萃取法处理血浆样品,以齐墩果酸为内标,采用ESI离子源,多重反应监测模式进行负离子检测,检测离子质荷比（m/z）为nodosin 361.0/256.9;齐墩果酸（内标）以m/z 455.2/407.2和m/z 455.2/455.2分别进行定性和定量。使用Diamonsil C18色谱柱（4.6 mm×250 mm,5μm）;以甲醇-5 mmol·L^-1乙酸铵水溶液为流动相,梯度洗脱;流速为1.0 m L·min^-1;柱温25℃。结果小鼠腹腔注射nodosin后,nodosin血药浓度在3.9-1000 ng·m L^-1范围内线性关系良好（r2〉0.99）,最低检测限为1.0 ng·m L^-1。日内、日间精密度（RSD）均小于9.8%,方法学考察均符合生物样品分析的要求。小鼠血浆中nodosin及内标物均无内源性基质的干扰。结论本研究建立的LC-MS/MS分析方法灵敏可靠,特异性高,适用于nodosin的药动学研究。Objective To develop a simplified,sensitive and specific liquid chromatography-tandem mass spectrometry（LC-MS/MS） assay for the determination of nodosin in mice plasma and then to investigate the pharmacokinetic profile of nodosin in mice with the method.Methods KM mice were intraperitoneally administered one-dose nodosin saline injection,and then blood samples were collected from the decapitated mice.Plasma concentrations were quantified by LC-MS/MS using oleanolic acid as the internal standard（IS） and using ESI ion source.Quantitation was performed using multiple reaction monitoring（MRM） mode in negative mode,and the optimized mass ion-pairs（m/z）361.0/256.9 and 455.2/407.2 were quantified for nodosin and IS respectively.After a liquid-liquid extraction pre-treatment for plasma sample,chromatographic separation was achieved on a reversed phase Diamonsil C18column（4.6 mm × 250 mm,5 μm）with mobile phase of methanol and 5 mmol·L-1ammonium acetate water solution by gradient elution.The flow rate was 1.0 m L·min^-1and the column temperature was set at 25 ℃.Results The linear range of the calibration curve was 3.9-1000 ng·m L^-1（r^2〉 0.99）.The lower limit of detection（LLOD） was 1.0 ng·m L^-1.The intra-day and inter-day precision was less than 9.8 %.No endogenous interference was found during the retention time for nodosin and IS.The method validation was subjected to the requirements of biological sample analysis.Conclusion The established method is sensitive,reliable and specific,and is appropriate for investigating the pharmacokinetics of nodosin.

关 键 词：NODOSIN 液相色谱-串联质谱法 药物动力学 

分 类 号：R285.5[医药卫生—中药学]