UPLC-MS/MS and multivariate analysis of inhibition of heterocyclic amine profiles by black pepper and piperine in roast beef patties

In this study, UPLC-MS/MS combined with principal component analysis (PCA) with the help of scores, loadings, and contribution plots was used to investigate the inhibition of the profiles of carcinogenic and mutagenic heterocyclic amines (HAs) in roast beef patties by different levels of black pepper (0.05, 0.10, and 0.15%, g/g) or piperine (0.005, 0.010, and 0.015%, g/g). Six HAs from three categories, imidazopyridine (PhIP), imidazoquinoxaline (IQx, MeIQx, and 4,8-DiMeIQx), and beta-carboline (harman and norharman), were detected and quantified in the roast beef patties. The PCA results demonstrated that both black pepper and piperine can significantly suppress the HA profiles and that lower levels of black pepper or piperine lead to better inhibition of HA formation, except the HA profiles of beef patties with 0.005% and 0.01% piperine were similar. In comparison, piperine inhibited HA formation better than did black pepper for all six HAs, as shown by the contribution plot. The total HAs were inhibited by all levels of black pepper (42%, 34%, 25%) and piperine (62%, 60%, 56%) in a dose-dependent manner, although black pepper suppressed only PhIP, IQx, MeIQx, and 4,8-DiMeIQx, whereas piperine inhibited all six HAs. The results show that UPLC-MS/MS in conjunction with PCA is a powerful tool for screening the inhibitory effects of spices and their components along with their relationship to HAs and can thus be used for safety control in food processing procedures.