Effects of amides from pungent spices on the free and protein-bound heterocyclic amine profiles of roast beef patties by UPLC-MS/MS and multivariate statistical analysis

Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), principal component analysis (PCA) and orthogonal projection to latent structure-discriminant analysis (OPLS-DA) were used to explore the effects of the amide components (capsaicin, sanshoamide, piperine) of pungent spices widely consumed in China (chili pepper, Sichuan pepper and black pepper) on the profiles of 17 heterocyclic amines (HAs) from seven categories in roast beef patties. Four groups of HAs were detected and quantified in both free and protein-bound states: imidazopyridines (DMIP, 1,5,6-TMIP), imidazoquinolines (MeIQ), imidazoquinoxalines (IQx, 4,8-MeIQx) and β-carbolines (norharman, harman). Notably, different amounts of added capsaicin, sanshoamide and piperine had significant inhibitory effects on free DMIP, MeIQx, 4,8-DiMeIQx and harman formation in roast beef patties. Additionally, all three amide components significantly inhibited the formation of protein-bound MeIQ, 4,8-DiMeIQx, harman, norharman and MeAαC. In general, amide ingredients inhibited the formation of β-carbolines and total HAs by approximately 70%. Furthermore, the addition of 0.005% and 0.010% capsaicin suppressed imidazopyridine formation by 44% and 35%, respectively, and 0.015% capsaicin inhibited imdazoquinoxaline formation by 33%. These findings shed light on the effects of amide ingredients on the formation of free and protein-bound HAs.

Keywords: Amide ingredients; Heterocyclic amines; Orthogonal projection to latent structure-discriminant analysis; Principal component analysis; Protein-bound; UPLC-MS/MS.