Effect Of Pungent Spices And Their Characteristic Ingredients On The Formation Of Heterocyclic Amines In Roast Beef Patties

Heterocyclic amines(HAs)are groups of carcinogenic and mutagenic compounds generated during the processing of protein-rich materials under high temperature.Its formation mechanism and inhibition pathway have been paid much attention.Nowdays,addition of natural antioxidants such as phenolic compounds and spices has been always used for the inhibition of HAs.However,the former studies of our research group have demonstrated that some phenolic antioxidants in spices can not inhibit the formation of HAs,but significantly enhance the formation of HAs.Thus,it can be inferred that there are non-phenolic compounds in spices that can suppress the formation of HAs.Therefore,three pungent spices including chilli pepper,Sichuan pepper and black pepper with non-phenolic compunds as the characteristic ingredients were selected and the inhibition on HAs of the spices and the amides ingredients were also investigated.Furthermore,the possible inhibition pathway was also discussed.Firstly,a qualitative and quantitative method for 17 HAs in roast beef patties by UPLC-MS/MS was developed,and applied to investigate the effects of chili pepper,Sichuan pepper and black pepper(0.5%,1.0%,1.5%)on HAs formation in roast beef patties(225°C,10 min/side).The influence of amide compounds(capsaicin,sanshoamides,piperine)on the formation of HAs was also examined.Proximate analysis was also conducted to discuss whether spices and the characteristic ingredients could affect the quality of beef patties along with the inhibition of HAs.Finally,the intermediates of HAs including phenylacetaldehyde,formaldehyde,acetaldehyde,acetaldehyde and 2,5-dimethyl pyrazine in chemical model systems were investigated to further illustrate the inhibition pathway of amides.The results show that there are six kinds of HAs in roast beef patties,such as PhIP,IQx,MeIQx,4,8-DiMeIQx,Harman and Norharman.The addition of pungent spices can significantly inhibit the total content of HAs(P < 0.05),but different amounts have different inhibition rates.The best inhibition on total HAs could be obtained by 0.5% chili pepper,1.0% Sichuan pepper and 0.5% black pepper.Chili pepper can significantly inhibit the formation of PhIP(P < 0.05),but enhance the formation of other 5 kinds of HAs when added more than 1.0%.Except Harman and Norharman,Sichuan pepper could significantly inhibit remaining 4 kinds of HAs(P < 0.05)and the inhibition rate of IQx were 100%.Black pepper has the similar influence to Sichuan pepper.0.5% black pepper can completely inhibit the formation of IQx,and the inhibition rate is 42% for total HAs.We also explored the effects of amide compounds of three pungent spices including capsaicin,sanshoamides,piperine on the formation of HAs.0.005% capsaicin in the system can make inhibition rate as high as 80%.The inhibition rate of HAs could be significantly increased(P < 0.05)when sanshoamides adding ratio is from 0.005% to 0.01%,but nosignificant changes(P > 0.05)up to 0.015%.Low doses of piperine can completely inhibit IQx and 4,8-DiMeIQx.With the increasing of adding ratio of piperine,it reduced the inhibition effect on HAs.The inhibition effects of characteristic ingredients of the material are largely consistent with various spices and better than the corresponding spices.Then,the result of the above analysis was further validated by chemical model systems.Capsaicin and piperine could significantly inhibit the formation of 6 HAs in chemical model systems,which were better than in beef patties.The results have demonstrated that the inhibition effects of pungent species could be related to the amide compounds.The influence of spices and amides on the quality of beef patties was also inveatigated.It was found that in addition to protein,there was no significant difference(P > 0.05)in the content of fat,moisture,pH,free amino acids,minerals,texture parameters between blank and adding spices groups.Capsaicin and piperine have significant inhibitory effect for phenylacetaldehyde which is the intermediates of PhIP in the simulation system(P < 0.05).Different amide compounds have different influence in quinoxaline's intermediates,such as formaldehyde,acetaldehyde,2,5-dimethyl pyrazine.Capsaicin has significant promoting effect(P < 0.05),on the contrary,piperine has significant inhibitory effect(P < 0.05)in formaldehyde,acetaldehyde,2,5-dimethyl pyrazine.This results coincide with the former research about pungent spices.It further verified that pungent spices can reduce the content of HAs in roasted beef patties because the amide compounds have an effect on Strecker degradation reaction.