应用超高效液相色谱-串联质谱法测定血清中全氟磺酸和全氟烷酸前体物质

The application of UPLC-MS/MS to detect precursors of perfluorooctanoic acid and perfluoroalkyl carboxylates in serum

目的 建立超高效液相色谱-串联三重四级杆质谱（UPLC-MS/MS）测定血清基质中15种全氟磺酸以及全氟烷酸前体物质的方法.方法 血清中的目标化合物与四丁基铵硫酸氢盐结合后,以甲基叔丁基醚有机溶剂进行离子对提取,将收集的上清液用氮气吹干后用0.25 ml 1∶1甲醇和水溶液复溶,再经过0.2μm尼龙滤膜净化,采用Waters ACQUITY^TM BEH C18色谱柱（50 mm×2.1 mm×1.7 mm）分离,在电喷雾负离子源（ESI-）和多反应监测模式（MRM）下进行测定.结果 在15种目标化合物中,全氟己基-2-次磷酸盐、全氟己基-全氟辛基-次膦酸盐、全氟辛基-2-次膦酸盐三种物质使用外标法定量,三个添加水平的回收率在41.01％ ～ 112.13％之间,RSD在1.80％～11.63％之间.其余的12种物质使用同位素内标法定量,两个添加水平的回收率在70.25％～127.51％之间,RSD均在1.23％ ～ 15.45％.15种待测物质的检出限在0.1～ 5.0 pg/ml之间,定量限在0.2～ 10.0 pg/ml之间.并且对10份人血清样品进行了目标化合物的检测,部分物质被检出,其中全氟-1-辛基磺酰胺、N-乙基全氟-1-辛基磺酰胺、N-甲基全氟-1-辛基磺酰胺乙酸、N-乙基全氟-1-辛基磺酰胺乙酸的浓度范围分别为＜ LOD ～0.94 pg/ml、＜LOD ～ 10.08 pg/ml、＜LOD ～6.74 pg/ml、＜LOD ～ 1.04 pg/ml.结论 本方法简单、快速,灵敏度和准确度高,满足研究全氟磺酸及全氟烷酸前体物质人体负荷水平的需要.

Objective We established a method of UPLC-MS/MS that was to detect fifteen precursors of perfluoroalkyl sulfonates （PFSA） and perfluoroalkyl carboxylates （PFCA） in serum.Methods Briefly,TBAS solution was added to sera,then the mixed solution was extracted with aliquots of MTBE.The MTBE aliquots were combined,evaporated to dryness under nitrogen,and reconsituted in 0.25 ml of methanol and water （1∶1）.Then the reconstituted solution through 0.2 μm nylon syringe filter was collected.Chromatographic separation was performed using a Waters ACQUITY^TM BEH ^18C column （50 mm × 2.1 mm × 1.7 mm）.Analyte quantitation was performed in the negative electrospray ionization mode and multiple reaction monitoring （MRM）.Results Three target substances,6：6PFPi,6：8PFPi,8：8PFPi,were externally confirmed by standard addition.Rates of recovery for these three chemicals were from 41.01% to 112.13% in two standard levels.And the relative standard deviations （RSD） were lower than 11.63% and higher than 1.80%.The other twelve substances were quantified with internal standard.Moreover in two standard levels,rate of recovery for these chemicals ranged from 70.25% to 127.51%.And RSD were more than 1.23% and less than 15.45%.And the corresponding limit of detection （LOD） and limit of quantitation （LOQ） for all target substances were 0.1-5.0 pg/ml and 0.2-10.0 pg/ml.Then we detected these target substances in ten different human serum samples.The levels of few substances were higher than LOD.And the ranges of FOSA-M,N-EtFOSA-M,N-MeFOSAA,N-EtFOSAA were respectively 〈 LOD-0.94 pg/ml,〈 LOD-10.08 pg/ml,〈 LOD-6.74 pg/ml,〈 LOD-1.04 pg/ml.Conclusion The method,with high sensitivity and accuracy,could meet the actual testing requirements.