尿样中神经性毒剂代谢产物的HPLC/Q-TOFMS/MS检测方法研究

Determination of Nerve Agent Metabolites in Urine by High Performance Liquid Chromatography/Quadrupole-Time of Flight Tandem Mass Spectrometry

建立了尿样中甲基膦酸单乙酯(EMPA)、甲基膦酸单异丙酯(IMPA)、甲基膦酸频哪基酯(PMPA)3种神经性毒剂代谢产物的HPLC/Q-TOFMS/MS检测方法。以StrataSi-1型固相萃取小柱对尿样中的3种神经性毒剂代谢产物进行分离,HPLC/Q-TOFESIMS/MS进行测定,内标法定量。该方法对EMPA、IMPA、PMPA的线性范围均为5～320μg/L,相关系数均不低于0.9974;EMPA、IMPA、PMPA的加标回收率分别为57%、98%、81%;检出限(S/N≥3)均为0.1μg/L,定量下限(S/N≥10)均为1μg/L。并将该方法应用于禁化武组织(OPCW)首次生物医学样品分析演练未知尿样的检测,结果满意。

A high performance liquid chromatography with quadrupole-time of flight tandem mass spectrometric（HPLC/Q-TOF MS/MS）method was developed for the determination of three nerve agent metabolites,e.g.ethyl methylphosphonic acid（EMPA）,isopropyl methylphosphonic acid（IMPA）and pinacolyl methylphosphonic acid（PMPA）in urine.The variable experimental conditions for sample preparation and instruments were optimized.The target compounds were separated on a Strata Si-1 solid phase extraction（SPE）cartridge and analyzed by HPLC/Q-TOF MS/MS under negative electrospray ionization（ESI）.An internal standard method was used for quantitation analysis.Under the optimal conditions,the standard curves of three nerve agent metabolites were linear over the concentration range of 5-320 μg/L with correlation coefficients not less than 0.997 4.The recoveries of EMPA,IMPA and PMPA were 57%,98% and 81%,respectively.The limits of detection（LOD,S/N≥3）were 0.1 μg/L and the limits of quantitation（LOQ,S/N≥10）were 1.0 μg/L for all metabolites.The method was applied in the determination of the unknown urine sample from first confidence building exercise for biomedical sample analysis of OPCW,and the results indicated that the method was simple,speedy,accurate and sensitive.