Novel sample treatment method for the determination of free (E)-4-hydroxy-2-nonenal in meat products by liquid chromatography/tandem mass spectrometry using 4-hydroxy-2-nonenal-d₃ as internal standard

Rationale (E)-4-Hydroxy-2-nonenal (HNE) is a reactive secondary product of lipid oxidation with biological significance. The analysis of HNE is a challenge due to its volatility and high activity. Developing sample preparation and analytical tools for the determination of free HNE is crucial for better understanding the actual level of free HNE in meat products. Methods Liquid nitrogen freezing, subzero-temperature extraction and derivatization were employed for meat sample treatment. Liquid chromatography/tandem mass spectrometry with electrospray ionization in negative ion mode was used for the determination of free HNE after isotope-coded derivatization. Results High repeatability and good recoveries with a limit of quantification as low as 0.25 pmol/g were found. Nineteen out of 24 samples, including chilled/processed meat products and meat-based instant foods, were found to contain free HNE with a range of 0.014-1.160 nmol/g. Conclusions The proposed method showed satisfactory reliability, sensitivity and accuracy. We believe that such a sample preparation strategy will provide a powerful tool for better understanding the actual level of free HNE in meat products.