Abstract

Per- and polyfluoroalkyl substances (PFAS) are synthetic organic compounds that over the past several years, have witnessed a dramatic increase in scientific attention. As PFAS are predominantly accumulated in plasma, monitoring individual burden levels in plasma are typically achieved via some combination of protein precipitation and/or solid phase extraction (SPE), either in online or offline modes. This work describes an updated PFAS extraction workflow, using 96-well plate technology and protein precipitation that is rapid, simple, inexpensive, and amenable for large cohort studies. In brief, plasma proteins were precipitated using methanol and the resulting centrifuged supernatant was directly analyzed using UHPLC-MS/MS. We monitored 51 PFAS, which were quantified via isotope dilution and the effectiveness of the method was demonstrated by using NIST blood-based Standard Reference Materials (SRMs). This method resulted in recoveries ranging between 70 and 89% for all analytes. The 96-well design exhibited low limits of detection and only required sample volumes of 100 µL, thus resulting in an amenable method for high-throughput plasma/serum PFAS screening.• PFAS were directly quantified in plasma and serum samples;• No SPE needed after protein precipitation;• SRMs can be used to validate PFAS measurement in plasma/serum.

Highlights

  • Background contamination and extraction efficiencypolyfluoroalkyl substances (PFAS) originating from solvents, labware, hardware and instrumentation has been previously noted, with efforts aimed at both minimizing (e.g., PFAS replacement kits) and validating their presence

  • Flaherty and co-workers [1] have described a simple method for measuring plasma-bound perfluorooctanoic acid (PFOA) using a 96-well format in three steps: (i) protein precipitation carried out in an Argonaut protein precipitation column, arrayed in a 96well plate format (Isolute, Argonaut), followed by (ii) repeated vacuum cycles using the extraction plate manifold to draw the crash solvent through the column and (iii) the eluate from each column were transferred to autosampler vials [1]

  • Calibration curves for all 51 PFAS are summarized in Table S4 and include analyte concentration ranges, regression equations, R2 values and limits of detection (LOD) and quantitation (LOQ)

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Summary

Method Article

A rapid and simple method to quantify per- and polyfluoroalkyl substances (PFAS) in plasma and serum using 96-well plates. We monitored 51 PFAS, which were quantified via isotope dilution and the effectiveness of the method was demonstrated by using NIST blood-based Standard Reference Materials (SRMs). This method resulted in recoveries ranging between 70 and 89% for all analytes. Subject Area: More specific subject area: Method name: Name and reference of original method: Resource availability: Chemistry Analytical Chemistry Direct analysis of PFAS in biological samples using 96-well plates and protein precipitation Flaherty, J. Equipment used with the method include: a Fisher microplate vortexer (120 V, ADV, Fisher Scientific), a Sorvall ST16R centrifuge (Thermo Scientific), and a Vanquish UHPLC (ultra-high pressure liquid chromatography) coupled to a TSQ Quantis triple quadrupole mass spectrometer (Thermo Scientific)

Background information and method applicability
Background contamination and extraction efficiency
Method validation
Findings
Conclusion
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