Dataset supporting the validation and application of an LC–MS/MS method for legacy and emerging PFAS in serum and whole blood
Description
This dataset contains raw and processed data supporting the article “Development and validation of a simplified LC–MS/MS method for the simultaneous determination of legacy and emerging per- and polyfluoroalkyl substances in serum and whole blood”. The dataset includes data generated during method validation and application, including linearity experiments, matrix effect evaluation, recovery assays and the analysis of real biological samples. Data are organized by biological matrix and include calibration responses, recovery values, matrix effect calculations and PFAS concentrations measured in anonymized serum and whole blood samples. Concentrations are expressed in ng mL⁻¹, while recovery and matrix effect values are expressed as percentages. This dataset provides additional supporting information to facilitate transparency and reproducibility of the analytical validation and application results.
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Steps to reproduce
A liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the determination of 20 legacy and emerging per- and polyfluoroalkyl substances (PFAS) in serum and whole blood. Blank matrices were first screened to confirm the absence or negligible presence of the target analytes. Commercial bovine fetal serum was used for serum validation, while goat whole blood was used for whole blood validation. For sample preparation, 200 µL of serum or whole blood were transferred into 1.5 mL polypropylene centrifuge tubes. For validation experiments, blank matrices were fortified with the target PFAS at the required concentration levels, and procedural internal standards were added to all samples. After 1 h of equilibration, acetonitrile was added to obtain a final volume of 400 µL. Samples were vortex-mixed for 30 s and centrifuged at 10,000 rpm for 10 min at 4 °C. Then, 150 µL of the supernatant were diluted 1:1 (v/v) with ultrapure water before LC–MS/MS analysis. Instrumental analysis was performed using an LC–MS/MS system operating in negative electrospray ionization mode. Chromatographic separation was carried out using a C18 analytical column and a binary gradient based on ammonium acetate aqueous and organic (acetonitrile) mobile phases. A delay column was installed between the pump and the autosampler to minimize potential interferences from system-derived PFAS background signals. Linearity was evaluated using calibration standards covering the range of 0.008–17 ng mL⁻¹. Each calibration level was analyzed in six replicates, identified in the dataset as A, B, C, D, E and F. Calibration curves were fitted using weighted (1/x) linear regression. Matrix effects were evaluated separately for serum and whole blood by comparing solvent-based and matrix-matched calibration curves. Solvent A, B and C correspond to three independent solvent-based calibration curves, whereas Matrix A, B and C correspond to three independent matrix-matched calibration curves. Matrix effects were calculated as ME (%) = (slope matrix-matched calibration / slope solvent-based calibration) × 100. Accuracy and precision were assessed through recovery experiments performed at nine fortification levels: 0.035, 0.07, 0.14, 0.35, 0.70, 1.4, 3.5, 7.0 and 14 ng mL⁻¹. Individual recovery values were calculated by comparing the measured concentration with the nominal fortified concentration and are reported for each replicate and matrix. The validated method was applied to 40 anonymized human serum samples and 40 anonymized human whole blood samples to assess its applicability to real biological matrices. These samples were obtained from independent study populations and were not paired samples from the same individuals. Therefore, concentration levels and detection frequencies were reported separately for each matrix and were not used for direct comparison between serum and whole blood.
Institutions
- Universidad de Las Palmas de Gran CanariaCanary Islands, Las Palmas de Gran Canaria