Isotope dilution HPLC-MS/MS for simultaneous quantification of acrylamide and 5-hydroxymethylfurfural (HMF) in thermally processed seafood

The objective of this study was to develop an accurate, robust and rapid HPLC-MS/MS-based method capable of quantifying acrylamide and 5-hydroxymethylfurfural (HMF) simultaneously. A Phenomenex Synergi Fusion-RP C18 column (50 mm × 2.5 mm, 2 μm) was used, and the MS/MS instrument was operated in multiple reaction monitoring mode. The isotope dilution method was used to correct for the matrix effect from samples. This method demonstrated low limits of quantification (2.12 ng/mL for acrylamide and 4.86 ng/mL for HMF) and excellent linearity (R2 > 0.999). This method also demonstrated excellent quantification accuracy, precision, and recovery (87-110%). Using the method developed, target analytes were quantified in 14 thermally-processed seafood samples. Resulting concentrations ranged from 5.58 to 50.35 μg/kg for acrylamide, and from 12.54 to 2276.44 μg/kg for HMF. The proposed isotope dilution HPLC-MS/MS method is a valid and rapid technique for simultaneous analysis of acrylamide and HMF in seafood.