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

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 (50mm×2.5mm, 2μm) 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 (2.12ng/mL for acrylamide and 4.86ng/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 50.35μg/kg for acrylamide, and from 12.54 to 2276.44μg/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.