液相色谱-三重四极杆质谱法同时检测生物检材中百草枯及其代谢物  

Liquid Chromatography-triple Quadrupole Mass Spectrometry Simultaneously Detecting Paraquat and Metabolites in Biological Samples

作　　者：孟宇珍 龚铎 李文越 文岩 樊哲瑜 张潮[1] 王涛 傅善林 尉志文[1] 贠克明[1] MENG Yuzhen;GONG Duo;LI Wenyue;WEN Yan;FAN Zheyu;ZHANG Chao;WANG Tao;FU Shanlin;WEI Zhiwen;YUN Keming(School of Forensic Medicine,Shanxi Medical University,Jinzhong 030600,Shanxi,China;Criminal Investigation Brigade,Wanbailin Branch of Taiyuan Public Security Bureau,Taiyuan 030001,China)

机构地区：[1]山西医科大学法医学院,山西晋中030600 [2]太原市公安局万柏林分局刑侦大队,太原030001

出　　处：《刑事技术》2022年第1期49-57,共9页Forensic Science and Technology

基　　金：国家自然科学基金(8207072359);国家重点研发计划(2017YEC0803504,2018YFC0807403);国家科技基础性工作专项(2015FY11400);科技强警基础工作专项(2019GABJC31)。

摘　　要：目的建立生物检材中同时检测百草枯(paraquat,PQ)及其主要代谢物单季铵盐(monoquat)、百草枯-单吡啶酮(paraquat-monopyridone,MP)、百草枯-联吡啶酮(paraquat-dipyridone,DP)、4-羧基-1-甲基吡啶盐(4-Carboxy-1-methylpyridinium ion,MINA)的液相色谱-串联质谱(LC-MS/MS)检测方法。方法以百草枯氘代内标(Paraquat-d8 Dichloride,PQ-D8)作为内标,检材样品调节pH后,经乙腈沉淀蛋白,使用不同色谱柱洗脱,在多反应监测模式下检测。结果百草枯PQ、monoquat、MP、DP、MINA的线性范围分别是5~800ng/mL、0.5~80ng/mL、5~800ng/mL、2.5~400ng/mL、2~320ng/mL(r均高于0.993),日内、日间精密度(RSDs)分别在5%~14%、3%~13%、3%~15%、5%~13%、2%~15%之间,准确度(RE)分别在91%~116%、80%~100%、80%~111%、85%~114%、91%~114%之间。生物样品处理后自动进样器上室温放置72h,各物质的准确度分别在90%~119%、56%~125%、60%~110%、78%~98%、83%~117%之间。结论本测定方法前处理过程简便,分离效果好,提取效率高,可使用本方法对疑似百草枯中毒的检材进行原体及代谢物的检测,为案件提供法律依据;本实验优化了课题组前期建立的生物样品中百草枯及monoquat、MP的检测方法,参考其案例结果,检测相应检材,对比分析,该检测方法在原体含量大幅下降时,痕量代谢物仍可检出。Objective To establish an LC-MS/MS method for determination of paraquat(PQ)and its main metabolites[monoquat,paraquat-monopyridone(MP),paraquat-dipyridone(DP)and 4-carboxy-1-methylpyridinium ion(MINA)]in biological samples.Methods The biological samples(blood and tissues for this assay)were added with Paraquat-d8 Dichloride(PQ-D;),the internal standard substance,successively having them adjusted of pH values and precipitated of proteins with acetonitrile so that their supernatants were cleansed with cyclohexane and consecutively centrifuged to receive the lower layer of acetonitrile.The received fluid was undergone with drying,re-dissolved into methanol,centrifuged and filtrated through membrane,thereafter having been brought to elution with different chromatographic columns of LC-MS/MS apparatus.For the analysis of mass spectrometry into the eluate from column separation,LC-MS/MS apparatus was set under multiple reaction monitoring(MRM)mode plus positive ionization.Results Linear ranges were present with PQ,monoquat,MP,DP and MINA at each individual’s 5-800 ng/mL,0.5-80 ng/mL,5-800 ng/mL,2.5-400 ng/mL and 2-320 ng/mL(all rendering an r higher than 0.993),with their intra-and inter-day precision(RSDs)being 5%~14%,3%~13%,3%~15%,5%~13%and 2%~15%plus the accuracy(RE)being 91%~116%,80%~100%,80%~111%,85%~114%and 91%~114%,respectively.Besides,if the processed samples were placed with the autosampler at room temperature for 72 hours,each analyte(in the order of PQ,monoquat,MP,DP and MINA)demonstrated its own accuracy of 90~119%,56%~125%,60%~110%,78%~98%,83%~117%,respectively.Conclusions This approach set up here is of simple pretreatment,good separation effect and high extraction efficiency,capable of detecting PQ and metabolites from suspected paraquat poisoning samples.As the optimized update from our team’s earlier attempt to detect paraquat,monoquat and MP in biological samples,this approach could also have detected the trace metabolites even if the PQ is greatly reduced of content lower than certain favorable c

关 键 词：法医毒物分析 百草枯 液相色谱-串联质谱法 百草枯代谢物 

分 类 号：D919.1[医药卫生—法医学]