乙腈沉淀-超高效液相色谱-串联质谱法快速测定2种尿苯代谢物  

Rapid Determination of Two Kinds of Metabolites in Urine by Acetonitrile Precipitation-Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry

作　　者：代文侠 张利明[1,2] 辜艳琴 王春民 夏瑜[1,2] DAI Wen-Xia;ZHANG Li-Ming;GU Yan-Qin;WANG Chun-Min;XIA Yu(Suzhou Center for Disease Control and Prevention,Suzhou 215137,China;iangsu Key Laboratory of Occupational Health,Suzhou 215137,China)

机构地区：[1]苏州市疾病预防控制中心,苏州215137 [2]江苏省职业卫生重点实验室,苏州215137

出　　处：《分析化学》2024年第11期1735-1744,共10页Chinese Journal of Analytical Chemistry

基　　金：2023年江苏省预防医学及血地寄防科研课题项目(No.Ym2023076);苏州市第四批重点学科项目(No.SZXK202117)资助。

摘　　要：建立了超高效液相色谱-串联质谱(UPLC-MS/MS)同时快速测定人体尿液中反-反式黏糠酸(Trans,trans-muconic acid, t,t-MA)和苯巯基尿酸(S-Phenylmercapturic acid, S-PMA)2种苯代谢物含量的方法。尿液样品经酸化处理后,再经乙腈沉淀去除蛋白,收集上清液,经过氮吹浓缩、溶剂复溶后进行检测分析。采用酸性甲酸铵溶液和甲醇溶液作为流动相进行梯度洗脱,BEH C_(18)色谱柱分离,在多反应监测模式(MRM)下,采用内标法进行定量分析。以96孔样品收集板为前处理容器,可以对大批量样本进行快速分析。t,t-MA和S-PMA的线性检测范围分别为20~2000μg/L和0.5~20.0μg/L,相关系数均为0.9998,检出限分别为1.2和0.06μg/L;在实际尿液中的加标回收率分别为90.6%~103.4%和87.3%~101.7%,相对标准偏差(RSD)分别为1.5%~3.5%和1.8%~4.2%。采用本方法检测41份苯接触岗位职工的尿样,t,t-MA和S-PMA的检出率分别为95%和78%。与固相萃取法相比,本方法具有操作简便、快速高效、灵敏度好和准确度高等优点,适用于大批量尿样中t,t-MA和S-PMA的快速测定。A method for rapid,simultaneous determination of two metabolites of benzene,trans,trans-muconic acid(t,t-MA)and S-phenylmercapturic acid(S-PMA),in human urine by ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was established.Urine sample was acidified and protein was removed by acetonitrile precipitation.The supernatant was concentrated by nitrogen blowing and redissolved by solvent,and then analyzed by UPLC-MS/MS.The compounds were separated on a BEH C_(18) column with acidic ammonium formate solution and methanol solution as mobile phases for gradient elution and then quantified in multiple reaction monitoring(MRM)mode by internal standard method.Combined with the 96-well sample collection plate as a pre-treatment vessel,large quantities of samples could be rapidly analyzed.The linear ranges for detection of t,t-MA and S-PMA were 20-2000μg/L and 0.5-20μg/L,respectively,and the correlation coefficients were 0.9998.The limits of detection were 1.2 and 0.06μg/L,respectively.The recoveries were 90.6%-103.4%and 87.3%-101.7%,and the relative standard deviations(RSDs)were in range of 1.5%-3.5%and 1.8%-4.2%,respectively.The method was applied to detection of 41 urine samples of employees exposed to benzene.The detection rates of t,t-MA and S-PMA were 95%and 78%,respectively.Compared with solid phase extraction method,this method showed many advantages such as simple operation,rapidity and high efficiency,high sensitivity and accuracy,which was suitable for rapid detection of t,t-MA and S-PMA in large quantities of urine samples.

关 键 词：乙腈沉淀法 超高效液相色谱-串联质谱 尿液 反-反式黏糠酸 苯巯基尿酸 

分 类 号：R446.12[医药卫生—诊断学] O657.63[医药卫生—临床医学]