液相色谱-荧光检测器法测定人体血清中16种多环芳烃

Determination of 16 polycyclic aromatic hydrocarbons in human serum by high-performance liquid chromatography coupled with fluorescence detector

目的建立高灵敏度的超高效液相色谱串联荧光检测器检测人体血清中16种痕量多环芳烃的分析方法。方法200μl血清样品,加入400μl正己烷,涡旋振荡后高速离心分离,取上层有机相,重复萃取2次,合并萃取液,氮吹至近干,再复溶于100μl乙腈中。经PAH C18色谱柱(5μm,4.6 mm×150 mm),在甲醇-水为流动相下梯度洗脱,超高效液相色谱串联荧光检测器测定。结果血清中的16种多环芳烃在0.20μg/L~5.00μg/L浓度内呈良好的线性关系,线性相关系数均>0.998。该方法,检出限为0.1μg/L;加标回收率为60.5%~96.0%,RSD为1.5%~7.6%。结论该方法操作简便、生物样本用量少、检出限低,适用于人群中多种多环芳烃的监测。

Objective This paper aims to establish a high sensitivity analytical method for the determination of 16 trace polycyclic aromatic hydrocarbons(PAHs)in human serum by using high performance liquid chromatography tandem fluorescence detector(HPLC-FLR).Methods A total of 400μl n-hexane was added to 200μl serum sample,vortexed and separated by high speed centrifugation.The upper organic phase was obtained for repeated twice extraction.The extracted liquid was evaporated by nitrogen to dryness and re-dissolved in acetonitrile solution.The chromatography was performed on a PAH Cig column(5μm,4.6 mm×150 mm)for gradient elution with methanol-water as mobile phase,and determined by ultra-high performance liquid chromatography with tandem fluorescence detector.Results The 16 PAHs in serum showed a good linearity in the concentration range of 0.20μg/L to 5.00μg/L,and the linear correlation coefficients(r)were all greater than 0.998.The detection limit of this method is 0.1μg/L.The recoveries were within 60.5%-96.0%,and relative standard deviations(RSDs)were within 1.5%-7.6%.Conclusion The method is easy to operate and has less biological samples and low detection limit,which is suitable for the monitoring of many PAHs in human.