LC-MS/MS Method for Per- and Polyfluoroalkyl Substances (PFAS) Analysis in Food Products, First Action 2025.10.

Abstract

Background: Per- and Polyfluoroalkyl Substances (PFAS) are synthetic compounds widely used due to their water, grease, and dirt repellent properties. They are persistent and resistant to degradation, therefore they can be found in the environment, the food chain, and in human fluids. The European Union (EU) has established limits of quantification (LOQs) for four PFAS in produce, seafood, meat, eggs, fish oil, milk, and ready-to-eat food for infants and young children and set maximum levels (MLs) for the same analytes in eggs, fish, and meat products.

Objective: The study describes a quantitative LC-MS/MS method for the determination of 57 PFAS in food. The 55 analytes are quantified by isotopic dilution while two of the compounds, namely Capstone A and Capstone B are quantified using 2-level standard addition. Seven of the compounds have only one transition, which need another technique for their confirmation in case of a positive result in a sample. The scope comprises analytes included in the AOAC INTERNATIONAL Standard Method Performance Requirements (SMPRs®) 2023.003, cited in the EU Recommendation 2022/1431, and analytes surveyed by service laboratories in Europe or mentioned in the literature.Representative samples from several food categories (baby food puree, milk-based infant formula (powder), full cream milk powder, fish meat and fish oil, whole egg, soluble coffee, pet food and edible offal) were validated. Targeted performance requirements for LOQ, recovery and precision were as defined per AOAC SMPR 2023.003.

Method: The protocol involves an acetonitrile/water extraction. For 55 PFAS analysis in all matrices, a solid phase extraction (SPE) method is applied. Detection is conducted by LC-MS/MS and isotopic dilution is applied for quantification. Confirmation for single-transition analysis is performed by LC-HRMS. For infant formula and baby food, the extract is diverted to two workflows, one for the 55 PFAS and one for Capstone A & Capstone B, where a diluted aliquot is analyzed by LC-MS/MS and quantified by the standard addition procedure.

Results: The method was single laboratory validated in the nine food matrices with results generally in agreement with the criteria of the AOAC SMPR 2023.003, in terms of LOQs, recovery and precision. LOQs were generally in the low ng/kg range, and recoveries typically fell within 70-120%, with acceptable repeatability and intermediate precision. The performance of the method was further demonstrated by analyzing 27 additional food matrices.

Conclusions: The method described covers a wide range of food matrices and analytes compares favorably with most of the requirements of the SMPR 2023.003, it contains as well a larger scope of analytes compared to the SMPR 2023.003, giving an advantage of searching other potential sources of PFAS contamination. Following independent scientific review, the AOAC Expert Review Panel for PFAS in Food Methods determined that the method met the requirements of AOAC SMPR 2023.003 and approved it for First Action status as AOAC Official Method 2025.10.

Highlights: This work presents a validated LC-MS/MS method for 57 PFAS across diverse food matrices, meeting or exceeding most of the AOAC SMPR 2023.003 performance requirements and demonstrating broad applicability for routine monitoring. The method includes additional PFAS of emerging interest and was approved for AOAC First Action status, designated AOAC Official Method 2025.10.