Fully Automated Sensitive Quantitation of PFAS in Seafood for Regulatory Screening Using Triple Quadrupole LC/MS

Significance of the Topic

PFAS are widely detected in seafood and pose potential health risks via dietary intake.

Regulatory bodies such as the U.S. FDA, EFSA, EURL POPs, and AOAC enforce stringent limits on PFAS levels in seafood matrices.

Conventional analysis relies on labor-intensive manual sample preparation (QuEChERS, SPE, evaporation), which can introduce errors and limit throughput.

Study Objectives and Overview

The study aimed to develop and validate a fully automated, sensitive, and high-throughput method for quantifying 73 PFAS compounds in shrimp.

Key goals included compliance with regulatory MDL and LOQ requirements and integration of sample preparation with LC/TQ analysis to enhance laboratory productivity and accuracy.

Methodology and Instrumentation

The workflow combined QuEChERS salting-out extraction, µSPE cleanup, and a dilute-and-shoot approach on a CTC PAL3 Series 2 RTC autosampler.

MassHunter software managed automated sample preparation in parallel with Agilent 6495D triple quadrupole LC/MS data acquisition.

Used instrumentation:

• CTC PAL3 Series 2 RTC autosampler with dual park stations and tools for extraction, cleanup, dilution, mixing, centrifugation, and cooling
• Agilent 1290 Infinity II LC system (high-speed pump, multicolumn thermostat)
• Agilent 6495D triple quadrupole mass spectrometer with AJS ESI source in negative ion mode
• ZORBAX RRHD Eclipse Plus C18 analytical (2.1 x 100 mm, 1.8 µm) and guard columns at 55 °C, 10 µL injection, mobile phases: 5 mM ammonium acetate/water (A) and methanol (B)

Results and Discussion

The automated workflow enabled concurrent sample preparation and LC/TQ analysis, reducing idle time and boosting throughput.

Method sensitivity met U.S. FDA MDL requirements (MDL ≤10 ng/kg for 28 regulated PFAS) and achieved LOQs (0.1–0.3 µg/kg) aligning with EU, EURL POPs, and AOAC criteria.

Calibration linearity for all 73 analytes showed R2 ≥0.99; surrogate recoveries ranged from 70% to 130%, and internal standard RSD was ≤20%.

Matrix-spiked QC recoveries spanned 65%–135%, with critical PFOS, PFNA, PFOA, and PFHxS recoveries at 80%–120%.

Precision was high: over 93% of compounds achieved repeatability (RSDr) ≤19%, and interbatch reproducibility (RSDR) ≤20% (≤12% for 30 regulated PFAS).

Benefits and Practical Applications

• Streamlined, fully automated sample preparation minimizes human error and labor demands
• High sensitivity and reproducibility support regulatory compliance for seafood PFAS screening
• Parallel processing of preparation and analysis enhances laboratory efficiency

Future Trends and Opportunities

• Extension to diverse food and environmental matrices with expanded PFAS panels
• Integration with real-time data processing and AI-driven quality control
• Miniaturization and green chemistry approaches to reduce solvent consumption
• Standardization of automated workflows across multi-site laboratories

Conclusion

The validated automated workflow combining CTC PAL3 and Agilent 6495D LC/TQ delivers accurate, precise, and high-throughput PFAS analysis in shrimp, meeting stringent regulatory requirements and enhancing laboratory productivity for routine seafood screening.

Reference

• Agilent Technologies. A Fully Automated Workflow for PFAS Analysis in Seafood for Regulatory Screening. Publication Number: 5994-8011EN, May 2025.