Quantitative PFAS Analysis in Cosmetics Using the CTC PAL3 Series 2 RTC Autosampler with the 6495D Triple Quadrupole LC/MS System

Significance of the topic

Per- and polyfluoroalkyl substances (PFAS) are persistent, bioaccumulative chemicals used in many consumer products, including cosmetics, to provide water resistance, film forming, and emulsifying properties. Regulatory pressure worldwide (EU cosmetic and POPs regulations, Korea MFDS, national bans and proposed prohibitions) is increasing the demand for sensitive, reproducible analytical methods to detect PFAS at trace levels in complex cosmetic matrices. Reliable PFAS testing supports product safety, regulatory compliance, and risk management across manufacturing and testing laboratories.

Objectives and study overview

This application note describes development and validation of a fully automated end-to-end workflow for quantitative analysis of 74 PFAS in cosmetic products (liquid foundations, lipsticks, mascaras). The objectives were to: automate calibration, extraction, micro‑SPE cleanup and LC/MS injection on a single robotic platform; achieve low method detection limits and validated limits of quantitation suitable for current regulatory thresholds; and demonstrate accuracy, precision, and throughput on real cosmetics.

Methodology

• Samples: Seven commercially available cosmetic products were evaluated. A representative liquid foundation with trace background PFAS served as QC matrix; others were treated as unknowns.
• Sample handling: 0.20 ± 0.01 g of sample was weighed into 10 mL vials (manual), then processed fully on the autosampler.
• Automated sample preparation steps (PAL3 Series 2 RTC): surrogate/internal spiking, addition of hexane as dispersant, addition of extraction solvent (MeOH/ACN 50:50), vortex mixing, centrifugation, micro‑SPE conditioning and loading, elution into injection vials, ISTD addition and direct LC injection.
• Calibration: Twelve automated calibration levels spanning 1–50,000 ng/L were prepared on‑platform; surrogates and ISTDs were included in every level.
• QC strategy: Matrix QC spikes at LSQ (2.5 µg/kg), MSQ (25 µg/kg), HSQ (250 µg/kg) in triplicate (three technical preps × two injections each). Procedural and matrix blanks were included to monitor contamination/background.
• Data processing: Compound identification and quantitation with Agilent MassHunter and Quantitative Analysis software; MDLs calculated from replicate QC standard deviation * 3.14; LOQvali established from QC performance and identification criteria.

Instrumentation used

• Autosampler / automation: CTC PAL3 Series 2 RTC autosampler equipped with multiple liquid syringe tools, dilutor, micro‑SPE tool, LC injection valve, tray cooler, centrifuge and related modules. All solvent tubing and consumables were PFAS‑tested.
• UHPLC: Agilent 1290 Infinity II UHPLC (high‑speed pump, multicolumn thermostat).
• Column: Agilent ZORBAX RRHD Eclipse Plus C18, 2.1 × 100 mm, 1.8 µm; column temperature 55 °C.
• Mass spectrometer: Agilent 6495D triple quadrupole LC/TQ with Agilent Jet Stream (AJS) ESI source operated in combined negative and positive polarities; unit resolution in Q1/Q3.
• Chromatographic/ion source conditions: 0.4 mL/min flow, mobile phases 5 mM ammonium acetate (A) and methanol (B), 2 µL injection, run time ≈ 14.5 min, sheath/aux gas and temperatures optimized for sensitivity.

Main results and discussion

• Scope and sensitivity: Of 74 target PFAS, 72 compounds had method detection limits (MDLs) < 1 µg/kg. Two FTCAs (6:2 FTCA, 8:2 FTCA) were not determined by LC/TQ in this matrix due to interference; the authors suggest GC/TQ as a complementary technique for volatile PFAS classes.
• Validated limits of quantitation (LOQvali): 64 analytes achieved an LOQvali of 2.5 µg/kg, including regulated compounds PFOA and PFOS—well below EU POPs unintentional trace contaminant thresholds (≤ 25 µg/kg).
• Recovery and precision: Matrix‑spiked recoveries across QC levels showed strong extraction efficiency. Over 82% of targets returned recoveries in the 65–135% range; regulated PFAS (PFHxS, PFOS, PFNA, PFOA, PFDA, tetraconazole) displayed LSQ recoveries of 86–115%. More than 90% of compounds had within‑batch RSD < 15% across QC levels. For key analytes, recoveries and RSDs were often tighter (recoveries ~89–115% and RSD < 5% reported for select PFAS).
• Matrix effects and exceptions: A small number of analytes exhibited elevated LOQs (e.g., PFBPA, FHxSA, PFDPA, FDSA) due to positive residues in the matrix blank. Six compounds showed poor recoveries (< 30%) in cosmetic matrices and were quantified at LLOQ in solvent instead.
• Contamination control: Solvent and procedural blanks showed negligible PFAS background compared with low‑spike QC, demonstrating effective mitigation of external contamination through PFAS‑tested consumables and a dedicated automated workflow.
• Real sample findings: Multiple PFAS classes (PFCA, PFECA, PFPA, PFSA, FASA, FASAA) were detected above MDLs in some mascaras and lipsticks, illustrating the workflow’s applicability for routine product screening.

Benefits and practical applications

• End‑to‑end automation reduces manual labor, human error, and idle instrument time by integrating calibration, extraction, cleanup and LC injection on one platform.
• Micro‑SPE on the autosampler minimizes hands‑on cleanup and supports consistent recoveries across complex cosmetic matrices.
• Sensitivity and reproducibility meet regulatory needs for trace PFAS screening and quantitation, enabling laboratories to support compliance testing and quality assurance.
• Parallel sample preparation and continuous LC/MS acquisition increase throughput for routine monitoring programs.

Future trends and potential uses

• Broader adoption of automated sample preparation platforms for PFAS analysis across consumer products and environmental matrices is likely as regulatory scrutiny increases.
• Complementary instrumentation (e.g., GC/TQ) and workflows will be required for certain PFAS subclasses (e.g., FTCAs) that suffer matrix interferences in LC/TQ methods.
• Ongoing improvements in low‑background consumables, source design, and chromatographic/ionization strategies will further lower MDLs and expand the number of reliably quantifiable PFAS in complex matrices.
• Integration with laboratory information management systems (LIMS) and expanded isotope dilution panels will enhance QA/QC, traceability, and robustness for regulatory testing programs.

Conclusions

The integrated CTC PAL3 Series 2 RTC autosampler coupled with the Agilent 6495D triple quadrupole LC/MS delivers a robust, high‑throughput, and reproducible workflow for quantitative PFAS analysis in cosmetics. The method achieved sub‑µg/kg MDLs for most targets, LOQvali of 2.5 µg/kg for the majority of analytes including key regulated PFAS, and acceptable recoveries and precision in challenging cosmetic matrices. Automation of calibration, extraction, micro‑SPE cleanup and injection reduces manual intervention and supports routine regulatory screening and laboratory efficiency. Complementary techniques may be required for specific PFAS classes impacted by matrix effects.

References

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