超高效液相色谱-四极杆/静电场轨道阱高分辨质谱联用同时检测人体尿液中苯、甲苯和二甲苯的6种代谢产物  

Simultaneous determination of six metabolites of benzene,toluene and xylene in human urine by ultra-high performance liquid chromatography coupled to Q-exactive high resolution mass spectrometry

作　　者：万玲利 郑丽辉[1] 潘绥 柯宗枝[1] WAN Ling-li;ZHENG Li-hui;PAN Sui;KE Zong-zhi(Occupational Health Department,Fujian Center for Prevention and Control of Occupational Diseases and Chemical Poisoning,Fuzhou Fujian,350001,China)

机构地区：[1]福建省职业病与化学中毒预防控制中心职业卫生科,福建福州350001

出　　处：《职业与健康》2021年第8期1033-1039,1043,共8页Occupation and Health

基　　金：福建省卫生计生青年科研课题(2018-02-14)。

摘　　要：目的建立同时检测人体尿液中苯、甲苯和二甲苯的6种代谢产物的超高效液相色谱-四极杆/静电场轨道阱高分辨质谱联用方法。方法尿液样品经固相萃取柱净化后,在最佳色谱条件下分离,采用Targeted-SIM/dd-MS2负离子模式扫描,一级精确母离子和二级碎片离子对样品快速定性筛查、一级精确母离子外标法定量分析。优化了色谱柱、流动相、柱温等色谱条件和样品前处理方法及质谱扫描模式等实验参数;测定了方法的检出限、准确度及加标回收等指标;用建立的方法检测了28家涉苯系物企业接触苯系物职工班后尿液中6种代谢产物。结果人体尿液中苯、甲苯、二甲苯的6种代谢产物尿中反-反式粘糠酸(tt-MA)、苯巯基尿酸(SPMA)、马尿酸(HA)、2-甲基马尿酸(2-MHA)、3-甲基马尿酸(3-MHA)和4-甲基马尿酸(4-MHA)在1.0~500μg/L(SPMA:0.1~50μg/L)范围内均有良好的线性关系,相关系数均大于0.999 8,检出限为0.001~0.050μg/L。实际样品的平均加标回收率在90.4%~104.4%范围内,相对标准偏差均低于4.2%。28家涉苯系物企业所采集的155份尿液样品中6种待测物均未超出职业接触限值。结论该方法前处理简单、灵敏度高、检出限低,可有效降低样品假阳性的可能性,适用于接触苯、甲苯和二甲苯生物样本监测指标的同时检测。Objective To establish a method for the simultaneous determination of six metabolites of benzene,toluene and xylene in human urine by ultra-high performance liquid chromatography coupled to Q-Exactive high resolution mass spectrometry(UHPLCQExactive HRMS). Methods The urine samples were purified by SPE column firstly,and then separated by chromatography under the optimal chromatographic conditions. Mass spectrometric detection was carried out in negative ionization modes by the experiment method of Targeted-SIM/dd-MS2 to conduct rapid qualitative screening of the sample by the precursor ion and the fragment ion,and quantitative analysis by external standard method with the precursor ion for characteristic ions. The chromatographic conditions such as chromatographic column,mobile phase,column temperature and some other experimental parameters such as sample pretreatment,mass spectrometry scanning mode were optimized. The detection limit,accuracy and standard recovery of the method were measured. Six metabolites in the urine of workers exposed to benzenes in 28 benzene-related enterprises were detected by the established method. Results Six metabolites of benzene,toluene and xylene in human urine such as urinary trans,trans-muconic acid(tt-MA),S-phenylmercaputuric acid(SPMA),hippuric acid(HA),2-methylhippuric acid(2-MHA),3-methylhippuric acid(3-MHA) and 4-methylhippuric acid(4-MHA) demonstrated good linear relationship with concentrations in the range of 1.0-500 μg/L(SPMA:0.1-50 μg/L). The correlation coefficients(r) were all higher than 0.999 8 and the limits of detection were in the range of 0.001-0.050 μg/L. The standard recovery rate of the actual samples of the six compounds were in the range of 90.4%-104.4% and the relative standard deviations were lower than 4.2%. None of the six testedsubstances exceeded the exposure limits in each one of the 155 samples from 28 benzene-related enterprises. Conclusion The developed method is a simple and sensitive analytical method with low detection limit.It also can ef

关 键 词：超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法 苯系物生物监测指标 反-反式粘糠酸 苯巯基尿酸 马尿酸 甲基马尿酸 尿液 固相萃取 

分 类 号：R115[医药卫生—公共卫生与预防医学]