Determination of PFAS in Tissue Using QuEChERSExtraction Followed by Noval Enhanced Matrix Removal Mixed-mode Passthrough Cleanup with LC-MS/MS Detection

Importance of the Topic

Per- and polyfluoroalkyl substances (PFAS) are persistent environmental contaminants that accumulate in organisms and pose health risks. Accurate determination of PFAS in tissues, such as fish and other biota, is critical for regulatory compliance, environmental monitoring, and safeguarding public health.

Objectives and Study Overview

This study aims to apply and validate a streamlined protocol combining QuEChERS extraction with enhanced matrix removal (EMR) passthrough cleanup for quantifying 40 PFAS in various tissue matrices. The goal is to meet EPA Method 1633 acceptance criteria while reducing preparation time, solvent usage, and contamination risks.

Sample Preparation Methodology

The procedure begins with 5 g of homogenized tissue mixed with water and acidified acetonitrile. QuEChERS extraction salts and ceramic homogenizers facilitate analyte partitioning. After centrifugation, the supernatant is diluted with water and loaded onto EMR PFAS cartridges. Gravity elution followed by low‐pressure flow yields cleaned extracts ready for analysis.

Used Instrumentation

• Liquid Chromatography: Agilent 1290 Infinity II with ZORBAX Eclipse Plus C18 column (2.1×100 mm, 1.8 µm) and InfinityLab PFC delay column for PFAS retention and separation.
• Mass Spectrometry: Agilent 6495D triple quadrupole (QqQ) operated in negative ion dMRM mode for sensitive and selective detection.

Main Results and Discussion

The QuEChERS+EMR approach reduced total sample preparation time by over 80% compared to the conventional EPA 1633 SPE workflow and cut solvent use by approximately 80%. Method performance showed high recovery (80–120%) across 40 PFAS, with repeatability (RSD) below 15% in eight tissue types (chicken, turkey, tuna, salmon, pork, tilapia, beef, cod). Matrix effects were effectively minimized, and chromatographic interference from cholic acids was resolved.

Benefits and Practical Applications

• Significantly shorter sample preparation (<4 h vs. 20–24 h) enhances laboratory throughput.
• Reduced solvent consumption aligns with green chemistry goals.
• Robust cleanup ensures reliable quantitation of long‐chain PFAS in complex tissues.

Future Trends and Opportunities

Advancements may include automation of the QuEChERS+EMR workflow, expansion to additional emerging PFAS analogs, and integration with high‐resolution MS for broader suspect screening. The method’s scalability supports large‐scale biomonitoring and regulatory surveillance.

Conclusion

The novel QuEChERS extraction coupled with EMR mixed‐mode passthrough cleanup offers a rapid, efficient, and validated solution for PFAS determination in tissues. It meets EPA 1633 criteria while improving productivity and analytical robustness.

References

1. U.S. Environmental Protection Agency. Method 1633, Revision A: Analysis of Per- and Polyfluoroalkyl Substances (PFAS) in Aqueous, Solid, Biosolids, and Tissue Samples by LC-MS/MS. EPA 820-R-24-007, December 2024.