AbstractThe inhibitory effects of vitamins (nicotinic acid, pyridoxamine [PM], and l‐ascorbic acid) and phenolic acids (ferulic acid and p‐coumaric acid) on the formation of 2‐amino‐3,8‐dimethylimidazo [4,5‐f] quinoxaline (MeIQx) were studied in a glycine/glucose/creatinine model system and fried tilapia cakes. The results showed that PM was the most potential inhibitor and the inhibition rates reached 82.72% and 78.54% in model system and fried tilapia cakes, respectively. Detailed formation mechanism of MeIQx was put forward to find the inevitable species in the non‐free radical formation mechanism of MeIQx. Dose‐dependent analysis of PM on methylglyoxal (MGO ) and MeIQx formation were studied by using model systems and the results showed that MGO and MeIQx were both reduced about 60% in reaction mixtures when the molar ratio of PM to glycine was 1:16, which indicated that MGO is a key intermediate on the pathway of MeIQx formation. Quantum chemistry calculations showed that PM can act as a useful inhibitor to inhibit the formation of MeIQx and react with MGO to form new compounds. A pathway for the inhibitory activity of PM against MeIQx formation was proposed.Practical ApplicationPyridoxamine was the most effective inhibitor against heterocyclic aromatic amines (HAAs) and could be applied to a variety of food systems. While the inhibitory mechanism is still unclear. Detailed formation mechanism of MeIQx was put forward first and suggested methylglyoxal as an inevitable species in the non‐free radical formation mechanism of MeIQx in this study. Pyridoxamine trapping methylglyoxal is likely a key mechanism against the generation of MeIQx was demonstrated by quantum chemistry calculation and experimental demonstration. These findings may provide effective suggestions for reducing HAAs and similar toxicants in daily cuisine.