固相支撑液液萃取结合超高效液相色谱-串联质谱法同时测定尿液中7种多环芳烃代谢物

建立了固相支撑液液萃取法(SLE)结合超高效液相色谱-串联质谱法(UPLC-MS/MS)测定尿液中1-羟基萘、2-羟基萘、2-羟基芴、9-羟基芴、2-羟基菲、9-羟基菲和1-羟基芘7种羟基多环芳烃(OH-PAHs)的方法。实验将0.8 mL尿液酶解液经SLE萃取,考察了洗脱溶剂种类及体积的影响,并对氮吹浓缩压力进行了优化。尿液样品使用6 mL乙酸乙酯洗脱,洗脱液在48 kPa(25℃)压力下氮吹至干后,经UPLC-MS/MS对7种OH-PAHs进行分析,采用同位素内标法定量。方法在0.3～3 000 mg·L^-1范围内线性关系良好(r≥0.995 1),检出限和定量下限分别为0.05～1.0 mg·L^-1和0.15～3.0 mg·L^-1。加标3个浓度水平的回收率为70.8%～117%,日内(n=3)和日间(n=3)相对标准偏差分别为3.7%～9.6%和3.7%～11%。采用本方法对10例石油工人和10例对照人群(在校大学生)尿液中的OH-PAHs水平进行分析,除2-羟基菲和9-羟基菲外,其余5种OH-PAHs在石油工人尿液中的含量均高于对照人群(p<0.05),差异具有统计学意义。该方法经上样和洗脱两步完成样品前处理,简单高效,可用于尿液样品中OH-PAHs的分析。

Simultaneous Determination of 7 Hydroxyl Polycyclic Aromatic Hydrocarbons in Urine by Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectrometry with Solid Supported Liquid-Liquid Extraction

An ultrahigh performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) with solid supported liquid-liquid extraction(SLE) was developed for the determination of seven hydroxyl polycyclic aromatic hydrocarbons(OH-PAHs) in urine samples.0.8 mL enzymatic hydrolyzed urine sample was extracted on a SLE cartridge.Parameters for SLE,including type and volume of eluting solvent,and nitrogen evaporation pressure were optimized.The adsorbed analytes were eluted with 6 mL ethyl acetate,and the collected eluate was evaporated to dryness under nitrogen at a pressure of 48 kPa at 25 ℃,then analyzed by UPLC-MS/MS and quantified by the internal standard method.Under the optimized conditions,the calibration curves for the analytes were linear in the range of 0.3-3 000 mg·L^-1,with correlation coefficients not less than 0.995 1.The detection limits(S/N=3) and quantitation limits(S/N=10) were in the ranges of 0.05-1.0 mg·L^-1 and 0.15-3.0 mg·L^-1,respectively.The recoveries for OH-PAHs in urine samples at three spiked levels ranged from 70.8% to 117%,with the intra-day(n=3) and inter-day(n=3) RSDs of 3.7%-9.6% and 3.7%-11%,respectively.The proposed method was successfully applied in the determination of OH-PAHs in urine from 10 petroleum workers and 10 control groups(college students).Except 2-hydroxy-phenanthrene and 9-hydroxy-phenanthrene,the concentrations of the other five OH-PAHs in the urine samples of petroleum workers were much higher than that in the control samples(p<0.05).The proposed SLE method only included two steps,therefore,the proposed method is applicable for the facile and efficient monitoring of OH-PAHs in urine samples.