生物样品中6种抗凝血鼠药的测定

Determination of 6 anticoagulant rodenticides in biological samples

目的：建立检测生物样品中抗凝血类鼠药（杀鼠灵、杀鼠醚、溴敌隆、大隆、敌鼠、氯敌鼠）的方法。方法：以高效液相色谱（HPLC）分离,紫外（UV）检测器定量检测,以高效液相色谱-串联质谱（HPLC-MS/MS）进行样品确认。通过对样品的预处理、色谱条件、质谱参数等的优化选择,采用MAX固相萃取柱对样品净化提纯,Phenom enex C18柱分离,紫外检测波长为285 nm。阳性样品采用子离子扫描（Daughter-scan）方式检测,将得到的特征离子作为鉴别鼠药的定性确认依据。结果：经过方法回收率、精密度等的方法学验证,杀鼠灵、杀鼠醚、溴敌隆、大隆、敌鼠和氯敌鼠的最低检出限分别为0.298、0.165、0.277、0.120、0.184、0.267μg/ml。线性范围为0.3～11μg/ml,线性相关系数r^2〉0.999,回收率在60.1%～106.5%之间。结论：该法简单快速,分离效果好,定性准确,能够同时对生物样品中多种抗凝血类鼠药进行检测,能够满足日常的检测需求。

Objective：To establish a method for determination of six anticoagulant rodenticides Warfarin,Coumatetralyl,Bromadiolone,Brodifacoum,Diphacinone,Chlorophacinone）in biological samples.Methods：Six anticoagulant rodenticides were determined by HPLC,and confirmed by HPLC-MS/MS.After optimization of sample preparation,separation condition and MS/MS parameters,a MAX solid phase extraction（SPE） column was selected to clean up samples,analytes were determined with UV detector（285 nm） after separated by a Phenomenex C18 column.Positive samples were confirmed by HPLC-MS/MS with Daughter-scan mode.Results：The limits of detection（LOD） were 0.298,0.165,0.277,0.120,0.184 and 0.267 μg/ml for Warfarin,Coumatetralyl,Bromadiolone,Brodifacoum,Diphacinone and Chlorophacinone,respectively.Linear range was from 0.3 to 11 μg/ml,with coefficient r^2〉0.999.The mean recoveries were between 60.1% and 106.5%.Conclusion：The method is simple and fast and has a good separation,so it can meet the need for determination of 6 anticoagulant rodenticides in biological samples in routine analysis.