超高效液相色谱-串联质谱法测定血清和尿液中有机磷酸酯及其代谢物  

Determination of Organophosphate Esters and Metabolites in Serum and Urine by Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

作　　者：吴文琪 王晓霞[3] 刘文彬 高丽荣[1,2,4] 于洋 贾天琪 石喆元 何蕴琛 邓静霖 陈春赐 WU Wen-Qi;WANG Xiao-Xia;LIU Wen-Bin;GAO Li-Rong;YU Yang;JIA Tian-Qi;SHI Zhe-Yuan;HE Yun-Chen;DENG Jing-Lin;CHEN Chun-Ci(State Key Laboratory of Environmental Chemistry and Ecotoxicology,Research Center for Eco-Environmental Science,Chinese Academy of Sciences,Beijing 100085,China;College ofResources and Environment,University of Chinese Academy of Sciences,Beijing 101408,China;Department of Endocrinology,Beijing Hospital,Beijing 100730,China;Hangzhou Institute for Advanced Study,Universityof Chinese Academy of Sciences,Hangzhou 310024,China)

机构地区：[1]中国科学院生态环境研究中心,环境化学与生态毒理学国家重点实验室,北京100085 [2]中国科学院大学资源与环境学院,北京101408 [3]北京医院内分泌科,北京100730 [4]国科大杭州高等研究院,杭州310024

出　　处：《分析化学》2024年第9期1346-1354,共9页Chinese Journal of Analytical Chemistry

基　　金：浙江省重点研发计划项目(No.2021C03176);国家重点研发计划项目(No.2022YFC3902302);国家自然科学基金项目(No.22076207)资助。

摘　　要：建立了一种通过少量人血清(1.0 mL)或尿液(1.5 mL)样本,同步分析血清和尿液中19种有机磷酸酯(OPEs)及其二酯代谢物(di-OPEs)的方法。采用乙腈进行液-液提取,ENVI-18固相萃取柱净化,通过超高效液相色谱-串联质谱联用仪(UPLC-MS/MS)对目标化合物进行测定。采用Shim-pack GIST C18色谱柱(100 mm×2.1 mm,2μm)和Shim-pack GIST-HP(G)C18保护柱分离目标物,质谱采用电喷雾离子源(ESI),以多反应监测模式(MRM)的正/负离子模式进行检测。本方法可在15 min内实现所有目标化合物的基线分离,目标化合物在2~100 ng/mL浓度范围内均具有良好的线性关系,血清基质和尿液基质的方法检出限分别为0.001~0.178 ng/mL和0.001~0.119 ng/mL,加标回收率为30.5%~126.8%,相对标准偏差(RSDs)为1%~23%。应用本方法对11个受试者配对的血清和尿液样本进行检测。在所有测试样品中,OPEs的内标回收率为61%~114%,di-OPEs的内标回收率为43%~103%。OPEs及di-OPEs在血清和尿液样本中表现出较高的检出率,血清样本中的OPEs和di-OPEs总浓度为1.580~3.843 ng/mL,尿液样本中的总浓度为5.149~17.537 ng/mL。研究结果表明,本方法能够有效检测生物样本中的OPEs及di-OPEs,同时也揭示了OPEs类化合物在人体内普遍存在的情况及其潜在的暴露风险。A new method was developed for simultaneous detection of total 19 kinds of organophosphate esters(OPEs)and their diester metabolites(di-OPEs)in human serum(1.0 mL)and urine(1.5 mL)with low volume of samples.The target compounds were determined using ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)after acetonitrile liquid-liquid extraction combined with purification using an ENVI-18 solid-phase extraction(SPE)column.OPEs and di-OPEs were separated using a Shim-pack GIST C18 column(100 mm×2.1 mm,2μm)with a Shim-pack GIST-HP(G)C18 guard column.An electrospray ionization source(ESI)was employed in mass spectrometry analysis,with positive/negative ion mode using the multiple reaction monitoring(MRM).All target compounds were separated within 15 min,and exhibited good linear relationshipsin the concentrationrange of2–100 ng/mL,with correlation coefficients(R2)above 0.994.Themethod detection limits(MDL)in serum ranged from 0.001 to 0.178 ng/mL and the MDL in urine ranged from 0.001 to 0.119 ng/mL.The recoveries of the analytes spiked in serum and urine matrices at two concentration levels were 30.5%–126.8%,with the relative standard deviations(RSDs)ranged from 1%to 23%.In addition,paired serum and urine samples from 11 patients were analyzed.For all samples tested,the internal standards of OPEs exhibited recoveries between 61%and 114%,whereas the internal standards for di-OPEs had recoveries ranging from 43%to 103%.OPEs and di-OPEs exhibited high detection frequencies in 22 serum and urine samples.Triethyl phosphate(TEP),tributyl phosphate(TBP),tris(2-ethylhexyl)phosphate(TEHP),tris(2-butoxyethyl)phosphate(TBEP),tris(1-chloro-2-propyl)phosphate(TCIPP),triphenyl phosphate(TPHP),tri-m-tolyl-phosphate(TMTP)and 2-ethylhexyl diphenyl phosphate(EHDPP)were universally detected in all serum samples.TCIPP was identified at the highest concentrations(median 0.548 ng/mL)in serum samples.In urine samples,the detection frequency for 12 kinds of target compounds reached 100%.Notably,TBP emerged

关 键 词：超高效液相色谱-串联质谱 有机磷酸酯 代谢产物 血清 尿液 

分 类 号：O657.63[理学—分析化学] R446.1[理学—化学]