Development and Validation of an LC-MS/MS Method for the Quantitation of 30 Legacy and Emerging per- and Polyfluoroalkyl Substances (PFASs) in Human Plasma, Including HFPO-DA, DONA, and CC6O4
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Abstract Per- and polyfluoroalkyl substances (PFASs) include persistent organic pollutants whose spread is still ubiquitous. Efforts to substitute substances of high concern with fluorinated alternatives, such as HFPO-DA (GenX), DONA (ADONA), and cC6O4, have been made. The aim of this work was to develop and validate an isotopic dilution liquid chromatography-tandem mass spectrometry (LC–MS/MS) method suitable to quantify 30 PFASs in human plasma. Analytes included legacy PFASs (PFOA, PFOS, and PFHxS), fluorinated alternatives (PFBA, PFBS, 6:2 FTSA, HFPO-DA, DONA, and cC6O4), and newly identified compounds (F-53B and PFECHS). The sample preparation was rapid and consisted of simple protein precipitation and centrifugation. Calibration standards and quality control solutions were prepared with a human pooled plasma containing relatively low background levels of the considered analytes. A complete validation was carried out: the lower limits of quantitation (LLOQs) ranged from 0.009 to 0.245 µg/L; suitable linearity (determination coefficients,R20.989–0.999), precision (2.0–19.5%, relative standard deviation), and accuracy (87.9–113.1% of theoretical) were obtained for considered concentration ranges. No significant variations of analyte responses were recorded under investigated storage conditions and during matrix effect tests. The external verification confirmed the accuracy of the method, although limited to 12 analytes. The method was also applied to 38 human plasma samples to confirm its applicability. The developed assay is suitable for large-scale analyses of a wide range of legacy and emerging PFASs in human plasma. To our knowledge, this is the first published method including cC6O4 for human biomonitoring. Graphical abstract
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Active efforts are currently underway to establish suitable methodologies for the detection of alternative PFAS compounds within human blood.
“Consequently, active efforts are currently underway to establish suitable methodologies for the detection of these alternative PFAS compounds within human blood [12].”
From The EU’s Per- and Polyfluoroalkyl Substances (PFAS) Ban: A Case of Policy over Science (2023) · cites this paper as [12]
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- Authors
- Frigerio, G., Cafagna, S., Polledri, E., Mercadante, R., & Fustinoni, S.
- Affiliations
- University of Milan (IT)
- Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico (IT)
- Corresponding author
- Fustinoni, Silvia
- Venue
- Analytical and Bioanalytical Chemistry
- Published
- 2021-12-15
- Volume
- 414
- Issue
- 3
- Pages
- 1259-1278
- ISSN
- 1618-2642 (ISSN-L)
- 1618-2650
- PMC
- PMC8760233
- OpenAlex
- W4200146031
- Cited by
- 80
- References
- 65
- Citations by year
- 2026: 23 · 2025: 22 · 2024: 10 · 2023: 23 · 2022: 2
- FWCI
- 3.30
- Type
- journal article
- Language
- en
- Update notices
- correction (2022-01-14)
- Open access
- hybrid
- doi.org
- Locations
- Analytical and Bioanalytical Chemistry (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Archivio Istituzionale della Ricerca (Universita Degli Studi Di Milano) (submittedVersion, other-oa)
- Funders
- Friedrich-Alexander-Universität Erlangen-Nürnberg
- Topics
- Physical Sciences › Environmental Science › Environmental Chemistry › Per- and polyfluoroalkyl substances research
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmaceutical Science › Fluorine in Organic Chemistry
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical chemistry methods development
- Keywords
- per- and polyfluoroalkyl substances, human plasma, HFPO-DA, GenX, Dona, perfluorooctanoic acid, PFOS, PFHxS, F-53B, human biomonitoring, isotopic dilution, method validation, matrix effects
- MeSH terms
- Chromatography, High Pressure Liquid, Environmental Monitoring, Environmental Pollutants, Fluorocarbons, Humans, Tandem Mass Spectrometry, Limit of Detection
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