漂浮固化分散液液微萃取/液相色谱法测定人体尿液中的6种羟基多环芳烃

Analysis of Six Hydroxylated Polycyclic Aromatic Hydrocarbons in Human Urine by Liquid Chromatography with Dispersive Liquid-Liquid Microextraction Based on Solidification of Floating Organic Droplet

建立了漂浮固化分散液液微萃取(DLLME-SFO)/液相色谱法(LC)同时测定人体尿液中6种羟基多环芳烃(OH-PAHs)的分析方法,并对萃取剂、分散剂、尿液p H值、盐浓度以及萃取时间等参数进行了优化。优化结果表明:尿液调节p H值为5.0,酶解后,加入20%氯化钠调节盐浓度,再加入40μL正十一醇作萃取剂和400μL丙酮作分散剂,涡旋后,3 000 r/min离心5 min,于-20℃冰箱中冷却10 min,待正十一醇凝固后,取出置于细胞瓶中,室温下溶解后,进液相色谱分析。在最优的条件下,该方法对6种OHPAHs的回收率为70.4%~129%,相对标准偏差为1.7%~14.8%;线性范围为2.5~150.0μg/L,相关系数(r2)为0.999 1~0.999 8;检出限和定量下限分别为0.5~1.0μg/L和1.5~3.0μg/L。该方法操作简单快速,可满足流行病学上大样本量尿液的检测需求。

A liquid chromatography with fluorescence detection(LC/FLD)combined with dispersive liquid-liquid microextraction based on solidification of floating organic droplet was developed for the simultaneous analysis of six hydroxylated polycyclic aromatic hydrocarbons(OH-PAHs)in human urine.The important parameters affecting the extraction efficiency,including the type and volume of extraction and dispersive solvents,urine pH value,salt concentration and extraction time were optimized.Under optimized conditions,the urine sample was adjusted to pH 5.0,then enzymatically hydrolyzed and added 20%NaCl to adjust the ion strength.400μL dispersive solvent acetone and 40μL extraction solvent 1-undecanol were rapidly injected into the urine sample.After vortexed and centrifuged at 3 000 r/min for 5 min,the urine was kept in a-20℃refrigerator for ten min untill the floating organic droplets were solidified and could be easily collected with a spoon into a clean cell bottle.The collected 1-undecanol was injected into LC for analysis after it melted.The results showed that there existed a linear range of 2.5-150.0μg/L for six targeted OH-PAHs in urine with their correlation coefficients(r 2)of 0.999 1-0.999 8.The extraction recoveries were in the range of 70.4%-129%with RSDs of 1.7%-14.8%.The limits of detection and quantification were in the ranges of 0.5-1.0μg/L and 1.5-3.0μg/L,respectively.With the advantages of simplicity and rapidness,the developed method could satisfy the requirments for large sample analysis on actual human urine.