色谱

色谱 ›› 2026, Vol. 44 ›› Issue (7): 764-776.DOI: 10.3724/SP.J.1123.2025.07008

• 研究论文 • 上一篇    下一篇

高通量固相萃取-超高效液相色谱-高分辨质谱法测定血清中29种全氟/多氟化合物和22种有机磷酸酯及其二酯代谢物

赵诗琪1,3, 丁皓4, 张许文琦4, 侯敏敏1,3,*(), 周婷婷4, 史亚利1,2,3, 蔡亚岐1,2,3   

  1. 1.国科大杭州高等研究院环境学院，全省新污染物环境与健康重点实验室，浙江 杭州 310024
    2.中国科学院生态环境研究中心，环境化学与生态毒理学国家重点实验室，北京 100085
    3.中国科学院大学，北京 100049
    4.浙江省生态环境科学设计研究院，全省新污染物环境与健康重点实验室，浙江 杭州 310007
  • 收稿日期:2025-07-23 出版日期:2026-07-08 发布日期:2026-07-09
  • 通讯作者: *E-mail：houminmin@ucas.ac.cn.
  • 基金资助:
    国家自然科学基金(22320102005);国家自然科学基金(42307481)

Determination of 29 per- and polyfluoroalkyl substances and 22 organophosphate esters and diester metabolites in human serum by high-throughput solid-phase extraction- ultra performance liquid chromatography- high resolution mass spectrometry

ZHAO Shiqi1,3, DING Hao4, ZHANG Xuwenqi4, HOU Minmin1,3,*(), ZHOU Tingting4, SHI Yali1,2,3, CAI Yaqi1,2,3   

  1. 1.Zhejiang Key Laboratory of Environment & Health of New Pollutants，School of Environment，Hangzhou Institute for Advanced Study，University of Chinese Academy of Sciences，Hangzhou 310024，China
    2.State Key Laboratory of Environment Chemistry and Ecotoxicology，Research Center for Eco-Environmental Sciences，Chinese Academy of Sciences，Beijing 100085，China
    3.University of Chinese Academy of Sciences，Beijing 100049，China
    4.Zhejiang Key Laboratory of Environment and Health of New Pollutants，Ecological and Environmental Science and Research Institute of Zhejiang Province，Hangzhou 310007，China
  • Received:2025-07-23 Online:2026-07-08 Published:2026-07-09
  • Supported by:
    National Natural Science Foundation of China(22320102005)

RichHTML

21

PDF

77

摘要：

监测血液中的污染物是评估其人体暴露水平和健康风险的重要依据。全氟/多氟化合物（PFAS）、有机磷酸酯（OPEs）及其二酯代谢物（di-OPEs）作为广泛共存于环境并具有显著毒性效应的污染物，对其在人体内暴露水平的监测具有重要意义。本研究基于96孔板固相萃取柱前处理方法，通过比较3种萃取柱，并对前处理步骤进行优化，建立了检测血清样品中29种PFAS、17种OPEs和5种di-OPEs的超高效液相色谱-高分辨质谱法（UPLC-HRMS）。使用Phree PLR 96孔板萃取净化，向其中依次加入300 µL含1%甲酸的乙腈，100 µL血清样品，最后用100 µL含1%甲酸的乙腈溶液洗脱。洗脱液真空浓缩后使用UPLC-HRMS测定目标化合物。PFAS和di-OPEs选用负离子模式，OPEs选用正离子模式，在全扫描/数据依赖的二级扫描模式（Full MS/ddMS2）下测定，使用内标法进行校正。结果表明，在优化条件下，目标物在0.05~50 ng/mL内具有良好的线性关系（R2>0.991），方法检出限（MDL）为0.000 120~0.274 ng/mL。此方法具有操作简单、萃取时间短、高通量等优点。应用该方法对2024年采集的济南地区15份血清样品进行测定，结果表明29种PFAS总量为6.71~379 ng/mL，其中PFOA中位含量为8.96 ng/mL，PFOS中位含量为4.07 ng/mL。17种OPEs总含量为0.015 0~10.5 ng/mL，中位含量为2.81 ng/mL。5种di-OPEs的总含量为<MDL~0.443 ng/mL，中位含量为0.015 0 ng/mL，说明这些污染物在人体血液中的复合暴露状况及其引起的潜在健康风险应该引起重视。

关键词: 全氟/多氟化合物, 有机磷酸酯, 96孔板, 超高效液相色谱-高分辨质谱, 血清

Abstract:

Monitoring pollutants in human blood is a crucial basis for assessing human exposure levels and health risks. Per- and polyfluoroalkyl substances （PFAS）， organophosphate esters （OPEs）， and their diester metabolites （di-OPEs） are widespread environmental co-contaminants with significant toxic effects， making it crucial to monitor their internal human exposure levels. However， existing studies have predominantly investigated these substances in isolation， lacking comprehensive research that simultaneously quantifies PFAS， OPEs， and di-OPEs in human serum. Based on the pretreatment method of 96-well solid phase extraction columns， this study compared three extraction columns and optimized the pretreatment steps to establish an ultra performance liquid chromatography-high resolution mass spectrometry （UPLC-HRMS） method for 29 PFAS， 17 OPEs， and 5 di-OPEs in human serum samples. Phree PLR 96-Wellplate was used as a cleanup plate. 300 µL of 1% formic acid in acetonitrile， 100 µL of the serum sample， and internal standards were added to the cleanup device in sequence. After standing for 5 minutes， the 96-well plate positive pressure device was used to press the mixture into the 96-well collection plate. Finally， the sample was eluted with 100 µL of 1% formic acid in acetonitrile. The eluate was collected and concentrated for the detection of PFAS， OPEs， and di-OPEs by UPLC-HRMS. PFAS and OPEs were detected using an Acclaim RSLC 120 C18 column， while di-OPEs were detected using an Acquity UPLC BEH C18 column. Both were subjected to gradient elution with methanol and 5 mmol/L ammonium acetate in water as the mobile phases. Sample ionization was performed using a heated electrospray ionization source （H-ESI）. PFAS and di-OPEs were analyzed in negative ion mode， while OPEs were analyzed in positive ion mode. Data acquisition was conducted in full-scan/data-dependent tandem mass spectrometry （Full MS/ddMS2） mode. Quantification was achieved using the internal standard calibration method to ensure measurement accuracy. The results showed that under the optimized conditions， the target compounds had good linear relationships in the range of 0.05–50 ng/mL （R2 > 0.99）， and the method detection limits （MDLs） of 29 PFAS， 17 OPEs and 5 di-OPEs were 0.000 120–0.274 ng/mL， 0.011 0–0.250 ng/mL， and 0.012 0–0.220 ng/mL， respectively， and the spiked recoveries were between 45.9% and 147.8%. The relative standard deviations （RSDs） were 1.2%–29.0%. Most PFAS and di-OPEs had matrix enhancement effects， and most OPEs had matrix inhibition effects. Among them， hexafluoropropylene oxide dimer acid （GenX） （196.5%） and trimethylphenyl phosphate （TMPP） （54.6%） had significant matrix enhancement and inhibition effects， respectively， which could be corrected with appropriate internal standards. The recoveries of these two substances after correction were 127.3% and 78.7%， respectively， which met the analysis requirements. The proposed approach offers significant practical benefits， combining straightforward operation with shortened extraction time and enhanced throughput， which was validated through analysis of 15 human serum samples collected in Jinan in 2024. The total contents of 29 PFAS were 6.71–379 ng/mL， with a median value of 22.9 ng/mL. Eight PFAS were detected with a detection frequency of 100.0%， with median contents of perfluorooctanoic acid （PFOA） and perfluorooctane sulfonate （PFOS） reaching 8.96 ng/mL and 4.07 ng/mL， respectively. The total contents of 17 OPEs were 0.015 0–10.5 ng/mL， with a median value of 2.81 ng/mL. The most frequently detected OPEs， with detection frequencies exceeding 60.0%， were triethyl phosphate （TEP）， tri-n-butyl phosphate （TnBP）， and triphenyl phosphate （TPHP）. The total contents of 5 di-OPEs were <MDL–0.443 ng/mL， with a median value of 0.015 0 ng/mL. Therefore， the combined exposure to these pollutants in human blood and its potential health risks demand serious attention.

Key words: per- and polyfluoroalkyl substances （PFAS）, organophosphate esters （OPEs）, 96-wellplate, ultra performance liquid chromatography-high resolution mass spectrometry （UPLC-HRMS）, serum

中图分类号: