人血清中有机磷酸三酯和二酯的生物监测方法的建立  

Ultra-High Performance Liquid Chromatography Coupled with Triple Quadrupole Mass Spectrometry for Simultaneous Determination of Organophosphate Triesters and Diesters in Serum for Human Biomonitoring Study

作　　者：顾俊婕 胡曼 张益宁 屈卫东[1,3] 周颖[1,2,4] GU Jun-Jie;HU Man;ZHANG Yi-Ning;QU Wei-Dong;ZHOU Ying(Centers for Water and Health,Key Laboratory of Public Health Safety,Ministry of Education,Fudan University,Shanghai 200032,China;Department of Chemistry in Public Health Science,School of Public Health,Fudan University,Shanghai 200032,China;Department of Environmental Health,School of Public Health,Fudan University,Shanghai 200032,China;Pudong New Area for Disease Control and Prevention,Fudan University Pudong Institute of Preventive Medicine,Shanghai 200136,China)

机构地区：[1]复旦大学公共卫生安全教育部重点实验室,公共卫生学院饮水与健康研究中心,上海200032 [2]复旦大学公共卫生学院营养与食品卫生学暨卫生化学教研室,上海200032 [3]复旦大学公共卫生学院环境卫生学教研室,上海200032 [4]复旦大学浦东预防医学研究院,上海200136

出　　处：《分析化学》2021年第8期1384-1392,I0009-I0012,共13页Chinese Journal of Analytical Chemistry

基　　金：国家重点研发计划项目(No.2017YFC1600500);国家自然科学基金项目(No.81373089)资助。

摘　　要：建立了超高效液相色谱/三重四极杆质谱联用技术测定血清中4种有机磷酸三酯(磷酸三苯酯(TPHP)、磷酸三乙酯(TEP)、磷酸三丁酯(TnBP)和磷酸三(2-氯乙基)酯(TCEP))和2种有机磷酸二酯(磷酸二苯酯(DPHP)和磷酸二乙酯(DEP))的生物监测分析方法。400μL样品经乙酸乙酯(1.2 mL)-甲酸(FA,60μL)萃取,采用冷冻真空离心进行浓缩,在Phenomenex Kinetex 2.6μm F5 LC色谱柱上分离,采用H_(2) O(含0.1%FA)-甲醇溶液梯度洗脱,ESI正负离子切换模式下多反应监测(MRM)扫描,同位素内标法进行定量分析。结果表明,6种目标物在较宽浓度范围内线性关系良好(R^(2)>0.99),检出限(LODs)为0.002-0.01 ng/mL,定量限(LOQs)为0.01-0.05 ng/mL,回收率为56.1%-112.2%,日内和日间精密度良好(RSD<15%)。采用本方法检测了30份上海居民血清样本,6种目标物的检出率为100%。本方法适用于大样本量人群血液中痕量有机磷酸三酯和二酯的生物监测研究,对于评价新型环境污染物有机磷酸酯(OPEs)的人群暴露与潜在的健康风险具有重要意义。A method for rapid and simultaneous determination of organophosphate triesters(triphenyl phosphate(TPHP),triethyl phosphate(TEP),tri-n-butylphosphate(TnBP),tri(2-chloroethyl)phosphate(TCEP))and diesters(diethyl phosphate(DEP)and diphenyl phosphate(DPHP))in human serum by ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry(UPLC-MS/MS)was developed.Serum sample(400μL)was extracted using 1.2 mL of ethyl acetate with 60μL of formic acid,followed by concentrating the extracts with freeze vacuum centrifugation technology.The sample was separated on a Phenomenex Kinetex 2.6μm F5 LC column with formic acid aqueous solution and methanol as the mobile phase,and then detected in turn under electrospray ionization(ESI)positive and negative ion multiple reactive monitoring(MRM)mode.Stable isotope-labeled internal standards were used to improve the assay performance of UPLC-MS/MS.The analytical parameters involving sample pretreatment,chromatographic separation and mass determination were optimized.Under the optimal conditions,low limits of detection(0.002-0.01 ng/mL),wide linearity(0.05-50 ng/mL)and good recovery(56.1%-112.2%)were achieved.The intra-day and inter-day precisions were expressed as variation coefficients within 15%.The proposed method was successfully applied to the determination of six target compounds in 30 serum specimens collected from residents living in Shanghai city.This method provided a reliable alternative for large-scale monitoring trace level of organophosphate triesters and diesters in human exposure measurement and risk assessment.

关 键 词：有机磷酸三酯 有机磷酸二酯 生物监测 超高效液相色谱-串联质谱 血清 

分 类 号：O657.63[理学—分析化学] R318[理学—化学]