2DLC-MS/MS法测定人尿液中多环芳烃暴露生物标志物  

Determination of PAHs metabolites in human urine by two-dimensional liquid chromatography-tandem mass spectrometry

作　　者：王天南 陈敏[1] 汪阳忠 李钢[1] 费婷[1] 吴达[1] 陆捷 WANG Tiannan;CHEN Min;WANG Yangzhong;LI Gang;FEI Ting;WU Da;LU Jie(Technology Center of Shanghai Tobacco Group Co.,Ltd.,Shanghai 201315,China)

机构地区：[1]上海烟草集团有限责任公司技术中心,上海201315

出　　处：《烟草科技》2022年第2期38-44,共7页Tobacco Science & Technology

摘　　要：为解决分析卷烟烟气多环芳烃(PAHs)暴露的生物标志物——羟基多环芳烃(OHPAHs)时基质干扰较大、前处理复杂等问题,建立了中心切割二维液相色谱-串联质谱(2DLC-MS/MS)同时测定人尿液中10种OHPAHs的方法。尿液样品仅需酶解过滤后直接上机,第一维色谱以水-甲醇体系为流动相,C;色谱柱分离除杂;第二维色谱以水-乙腈体系为流动相,PAH色谱柱梯度洗脱,多反应监测模式(MRM)下进行MS/MS分析;两维间通过在线稀释将目标流份切割至捕集柱保留。结果表明:(1)10种OHPAHs均达到基线分离。(2)方法的检出限和定量限分别为0.001~0.021、0.005~0.070 ng/mL,回收率为87.9%~129.1%,日内和日间RSD在5%以内。(3)该方法成功应用于6名吸烟者和6名非吸烟者的尿液分析。该方法前处理简单便捷,仪器全自动,有效解决了5种羟基菲同分异构体准确定量的难题,可为烟气PAHs暴露标志物测定提供更为高效、准确的技术手段。To simplify sample pretreatment and reduce interference caused by matrix components when analyzing hydroxylated polycyclic aromatic hydrocarbons(OHPAHs), which are recognized biomarkers of polycyclic aromatic hydrocarbons(PAHs) for cigarette smokers, a new method for simultaneously determining 10 OHPAHs in human urines was developed based on two-dimensional liquid chromatography-tandem mass spectrometry system(2DLC-MS/MS). The urine samples were directly analyzed after enzymatic hydrolysis of PAHs. The analytes were separated on a C;column with water-methanol as the mobile phases in the first chromatograph separation, cut to the trap column by online dilution, and then reserved and enriched on the column. Finally the analytes were separated by gradient elution on the PAH column with water-acetonitrile as the mobile phases in the second chromatograph. MS/MS analysis was performed in multiple reaction monitoring(MRM)mode. The results showed that:1) Baseline separation was realized for all 10 target OHPAH compounds. 2) The detection limits of the method for OHPAHs were 0.001-0.021 ng/mL and quantification limits were 0.005-0.070 ng/mL. The spiked recoveries ranged from 87.9% to 129.1%with intra-and inter-RSDs less than 5%. 3)The method was successfully applied to the analysis of the urines from 6 smokers and 6 non-smokers. This method achieved accurate quantitation of 5 hydroxyphenanthrene isomers with simple sample pretreatment and highly automated procedures, thus providing a more efficient and accurate technique for the determination of PAHs in tobacco smokers.

关 键 词：尿液 多环芳烃 羟基多环芳烃 生物标志物 中心切割 二维液相色谱-串联质谱 

分 类 号：TS416[农业科学—烟草工业]