同时定量检测水胺硫磷和水胺氧磷的液相色谱-串联质谱法及应用  

A liquid chromatography/tandem mass spectrometry based method for simultaneous determination of isocarbophos and isocarbophos oxon in rat blood

作　　者：魏霞[1] 钟玉环[1,2] 原梅[1] 谢剑炜[1] 李桦[1] 

机构地区：[1]北京军事医学科学院毒物药物研究所,100850 [2]烟台烟台大学药学院,264005

出　　处：《国际药学研究杂志》2012年第2期162-166,共5页Journal of International Pharmaceutical Research

基　　金：国家“重大新药创制”科技重大专项(2008ZXJ09006-001)

摘　　要：目的建立同时测定大鼠全血中水胺硫磷及其活性代谢产物水胺氧磷的液相色谱-串联质谱（LC-MS/MS）分析方法,用于毒代动力学研究中血药浓度的测定。方法采集大鼠全血样品后即刻用4倍体积冰冷乙腈沉淀蛋白。待测物用Zorbax SB C18柱（2.1 mm×50 mm,1.8μm）分离,以含有0.1%甲酸的水和甲醇为流动相系统进行梯度洗脱。采用ESI源正离子多反应监测模式分析,水胺硫磷、水胺氧磷和内标三唑磷的定量离子对分别为m/z 231→121、274→215和314→162。结果水胺硫磷和水胺氧磷在5μg/L～5 mg/L浓度范围内线性良好,最低定量限为5μg/L,回收率在63.1%～77.8%的范围内,准确度和精密度符合生物样品的检测要求。大鼠静脉注射3 mg/kg水胺硫磷后,水胺硫磷在血中很快下降,消除半衰期为22.2 min。活性代谢产物水胺氧磷的生成也较快,10～15 min达到（257.91±42.00）ng/ml的平均峰值,消除半衰期为17.6 min。水胺硫磷和水胺氧磷的AUC0~t分别为（528.62±49.15）和（191.11±33.23）h.μg/L。结论本研究首次建立了同时定量测定大鼠全血中水胺硫磷和水胺氧磷的LC-MS/MS分析方法,方法的专属性好、灵敏度高,适用于水胺硫磷及水胺氧磷在大鼠体内的毒物代谢动力学研究。Objective To develop and validate a liquid chromatography-tandem mass spectrometry method for simultaneous quantification of isocarbophos and its active metabolite isocarbophos oxon in rat blood.Methods Rat blood samples were treated with pre-cold acetonitrile（1∶ 4 in volume） for precipitating proteins.Chromatographic separation of isocarbophos and isocarbophos oxon was achieved on a Zorbax SB C 18 column（2.1 mm×50 mm,1.8 μm）by a gradient elution with water and methanol containing 0.1% formic acid.The MS detection was performed using an Agilent 6430 tandem mass spectrometer in positive ion electrospray and multiple reaction monitoring（MRM） mode.The ion transitions monitored were m/z 231→121,274→215 and 314→162 for isocarbophos,isocarbophos oxon and trizophos（the internal standard）,respectively.Results A good linearity was obtained over the concentration range of 5 μg/L-5 mg/L for both analytes,and the lower limit of quantification was 5μg/L.The recoveries of isocarbophos and its metabolite were all in the range from 63.1% to 77.8%.The accuracy and precision of the method met the requirement for bioanalysis of blood samples.The method was successfully used for a toxicokinetic study in rats.After an intravenous dose of 3 mg/kg,the blood concentration of isocarbophos declined rapidly with the elimination half-life of 22.2 min.The formation of isocarbophos oxon in rat blood was also rapid.The mean peak level of（257.91±42.00） ng/ml was reached in 10 ~ 15 min post-dose and its elimination half-life was 17.6 min.The AUC 0~t values for isocarbophos and isocarbophos oxon were（528.62 ± 49.15） and（191.11 ± 33.23）h.μg/L respectively.Conclusion The method is proved to be simple and sensitive,and suitable for toxicokinetic studies of isocarbophos.

关 键 词：水胺硫磷 水胺氧磷 液相色谱-串联质谱法 毒物代谢动力学 

分 类 号：R965[医药卫生—药理学]