在线固相萃取-高效液相色谱法同时测定人尿液中7种多环芳烃代谢物

Simultaneous Determination of Seven Metabolites of Polycyclic Aromatic Hydrocarbons in Human Urine by Online Solid Phase Extraction-High Performance Liquid Chromatography

采用双三元液相色谱系统结合荧光检测器,建立了在线固相萃取-液相色谱法同时测定人尿液中7种多环芳烃代谢物的方法。目标化合物首先在Turboflow Cyclone固相萃取柱上在线富集浓缩,然后通过六通阀转移至Hypersil Green PAH色谱柱,以乙腈-水为流动相进行梯度洗脱分离,流速1.0 mL/min,柱温30℃,荧光检测器检测,分离周期为20 min。在优化的色谱条件下,5～2000 ng/L或50～20000 ng/L范围内,7种多环芳烃代谢物均呈良好的线性关系（r≥0.999）,方法检出限为0.5～15 ng/L,加标回收率为80.7％～110.7％。应用本方法对吸烟和非吸烟人群尿液中7种多环芳烃代谢物的含量进行了测定,吸烟者尿液中的2-羟基萘、1-羟基萘、2-羟基菲、2-羟基芴、4-羟基菲、6-羟基显著高于非吸烟者。

A novel method for the simultaneous quantification of seven metabolites of polycyclic aromatic hydrocarbon in human urine was developed using online solid phase extraction-HPLC with double ternary liquid chromatography system combined with fluorescence detector. The target compounds were online concentrated on the Turboflow Cyclone solid phase extraction column at first, then transferred by the six-way valve to the Hypersil Green PAH column for separation with acetonitrile and water as mobile phase at a flow rate of 1. 0 mL/min and at 35 ℃. A single sample analysis cycle took only 20 min. Under the optimized chromatographic conditions, the method showed good linear relationship （ r≥0. 999 ） in the range of 5-2000 ng/L or 50-20000 ng/L. The LODs were 0. 5-15 ng/L, and the recoveries were 80. 7%-110. 7%. The proposed method was successfully applied in the detection of metabolites of polycyclic aromatic hydrocarbons in urine from several smokers and non-smokers. The concentrations of 2-hydroxynaphthalene, 1-hydroxynaphthalene, 2-hydroxyfluorene, 2-hydroxyphenanthrene, 4-hydroxyphenanthrene and 6-hydroxychrysene in the smokers urine were much higher than that in non-smokers.