Analysis of Per- and Polyfluoroalkyl Substances (PFAS) in Edible Fish Tissue

Importance of the Topic

PFAS contamination in food, particularly edible fish, poses health risks due to their persistence and bioaccumulation. Regulatory bodies now set strict limits for PFAS such as PFOS, PFOA, PFNA and PFHxS in fish tissues to protect human health.

Objectives and Study Overview

This study aimed to develop and demonstrate a robust analytical workflow for quantifying 25 PFAS compounds in various fish species. The approach integrates effective lipid removal, optimized LC/MS/MS parameters, and contamination control measures to ensure low detection limits and regulatory compliance.

Methodology and Instrumentation

The sample preparation employs Captiva EMR-Lipid cartridges for efficient removal of fats and lipids without analyte loss. Key steps include tissue homogenization with formic acid, acetonitrile extraction, centrifugation, and SPE cleanup.

• Agilent 1290 Infinity II UHPLC system equipped with a PFC-free HPLC conversion kit and a delay column to minimize PFAS background.
• Agilent ZORBAX RRHD Eclipse Plus C18 analytical column (2.1 x 100 mm, 1.8 μm) with a guard column and Quick Change inline filter.
• Agilent 6470B triple quadrupole LC/MS with Jet Stream ESI source, operated in negative ion mode.

LC conditions feature a gradient from 100% aqueous ammonium acetate to high organic composition, flow rate 0.3 mL/min, injection volume 20 μL, and column temperature at 50 °C. MS parameters include drying gas (230 °C, 4 L/min), sheath gas (250 °C, 12 L/min), nebulizer at 15 psi, and capillary voltage of 2500 V.

Key Results and Discussion

The workflow achieved consistent recovery of 25 PFAS analytes across 25 fish species with minimal matrix interferences. Lipid removal reduced ion suppression, enhancing method sensitivity and reproducibility. PFAS-free HPLC components significantly lowered background signals, enabling reliable detection well below regulatory thresholds.

Benefits and Practical Applications

• Compliance with EU Regulation 2022/2388 and other guidelines for PFAS in food.
• High throughput screening for food safety laboratories and environmental monitoring.
• Scalable approach adaptable to different fish matrices and other biological tissues.
• Improved data quality through rigorous contamination control.

Future Trends and Opportunities

Expanding compound coverage to emerging PFAS and transformation products using high-resolution MS. Automated sample preparation and integration with laboratory information management systems will increase throughput. Ongoing method validation for additional matrices such as shellfish, dairy, and plant-based foods can broaden monitoring capabilities.

Conclusion

The developed workflow provides a reliable, sensitive, and compliant solution for PFAS analysis in edible fish tissues. By combining effective lipid cleanup, PFAS-free instrumentation, and optimized LC/MS/MS settings, laboratories can achieve regulatory compliance and generate reproducible data for food safety and environmental studies.

Reference

1. Food Safety Magazine. EU Sets Limits for PFAS in Certain Foods. 2022.
2. Regulation (EU) 2022/2388, Official Journal of the European Union. 2022.
3. ACS Publications. Toxic PFAS found in freshwater fish. Environmental Science & Technology. 2023;101(3).
4. Pulster EL, Giardina M. Analysis of Per and Polyfluoroalkyl substances in edible fish tissue using Captiva EMR-Lipid and LC/MS/MS. Agilent Technologies Application Note 5994-5227EN. 2022.
5. Frontiers in Marine Science. Assessing per- and polyfluoroalkyl substances in sediments and fishes in a large, urbanized estuary. 2022;9:1046667.
6. Kamuf M et al. Reduce PFAS Background with Agilent PFC-free HPLC conversion kit. Agilent Technologies Application Note 5994-2291EN. 2021.