A rapid and simple method to quantify per- and polyfluoroalkyl substances (PFAS) in plasma and serum using 96-well plates

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.

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

“…The main strength of the present method is the quantitation of several emerging PFASs, with great sensitivity: LLOQs of the present work were lower for HFPO-DA, F-53B-related analytes, and DONA [16,43,44,56] and comparable for PFECHS [43] by comparing them with those reported by published methods. To the best of our knowledge, this is the first method able to quantify cC6O4 in human blood matrices.…”

“…Before injection, plasma and serum samples are usually treated to remove proteins [41][42][43][44][45] and/or purified with solid-phase extraction (SPE) [16,[46][47][48][49][50][51][52][53][54][55][56][57][58]. However, only a few methods considered the emerging compounds HFPO-DA [16,43,44], F-53B 1 3 [16,53,56], DONA [16,43], or PFECHS [43,45]. Also, we did not find in the scientific literature methods determining cC6O4 in human blood matrices.…”

“…Nevertheless, after performing some experiments, we decided to avoid its use for some reasons: (1) a matrix effect test suggested that the usage of SPE did not improve the burden of the matrix effect for the analytes and that the additional manual steps required affected reproducibility; (2) considerable contamination with PFBA derived from WAX cartridges was observed, as declared by the producer [80]; and (3) one of the considered analytes is not a strong acidic compound (PFOSA), thus requiring the collection and analysis of the eluate derived from the cartridge washing step, in turn reducing the throughput of the assay. Only a few other methods analysed PFASs with a sample preparation consisting only of a protein precipitation without further cleaning [42][43][44]81]. The main advantages are the few steps and short time required for sample preparation, the low amount of solvent used, and the lower cost for consumables.…”

“…Several methods have been developed for the quantitation of different chemical classes of PFASs in human serum and plasma, by implementing liquid chromatography coupled with tandem mass spectrometry as a suitable analytical technique for high sensitivity and selectivity. Before injection, plasma and serum samples are usually treated to remove proteins [41][42][43][44][45] and/or purified with solid-phase extraction (SPE) [16,[46][47][48][49][50][51][52][53][54][55][56][57][58]. However, only a few methods considered the emerging compounds HFPO-DA [16,43,44], F-53B 1 3 [16,53,56], DONA [16,43], or PFECHS [43,45].…”

A FAIR approach for detecting and sharing PFAS hot-spot areas and water systems

“…Per-and polyfluoroalkyl substances (PFAS) are a large group of 9000 persistent fluorinated synthetic chemicals that have been used since the 1950's and are detected in the environment and humans globally now [1][2][3]. PFAS sources are associated with various potential PFAS "users" that currently use or have used products containing PFAS.…”