Preliminary Study On The Substance And Mechanism Of Theinhibition Of Spice On The Formation Of ?-carbolines Heterocyclic Amines In Braised Meat

This study focused on the substance basis and mechanism of the inhibition of spices on the formation of ?-carboline heterocyclic aromatic amines(HAAs)in braised meat.Firstly,contents of HAAs in braised meat were determined by Centrifugal-SPE-HPLC method by optimizing the pretreatment method.Then,the effects of 8 spices on the formation of?-carboline HAAs in braised meat and the correlation between antioxidant activity and the inhibitory ability were fitted.Different ?-carboline HAAs chemical models were set up to filter the appropriate chemical system.The influence of different temperature and heating time on the formation of ?-carboline HAAs were studied.The main polyphenol components in two spices with good inhibitory effect were identified by LC-MS,and the dose-effect relationship of different polyphenols on the formation of ?-carboline in the chemical system were analyzed.Finally,the inhibitory mechanism of polyphenols on?-carboline HAAs was studied by studying the inhibitory and eliminate effect of polyphenols on ?-carboline HAAs,intermediates and carbonyl compounds.The main findings are as follows:(1)Determination of ?-carboline HAAs in Braised Meat by SPE-HPLCA method was optimized for the determination of ?-Carboline HAAs in sauce braised meat products by solid phase extraction-high performance liquid chromatography.The sample was extracted with dichloromethane,purified by centrifugation and a MCX SPE column,and then detected by a fluorescence detector.The results showed that the correlation coefficients of both HAAs were above 0.999 and the limits of detection were in the range from 0.17~0.30 ng/g.The recoveries of the two HAs spiked in chicken samples were 81.92%~93.37% with the relative standard deviations(RSD)between3.66% and 7.49%.The method is simple,sensitive and effective,and can meet the requirements of actual sample analysis.1-methyl-9H-pyrido[4,3-b]indole(Harman)and9H-pyrido[4,3-b]indole(Norharman)were detected in commercially available braised meat.The total amount of ?-Carboline HAAs in braised meat and marinade increased with the times of cooking.(2)Traditional Chinese spice and its effect on the formation of ?-Carboline HAAs in braised meatThe effects of eight spices on the formation of ?-Carboline HAAs were studied.Eight spices had different effects on ?-Carboline HAAs formation.Among them,dried ginger,green pepper and cinnamon had the strongest inhibitory effects,with the inhibition rate of above 40% at 5% of addition.The inhibition effect of tangerine peel and black pepper were relatively weak,under the 5% addition amount,the content of ?-Carboline HAAs decreased by 32.40% and 30.96%,respectively,and the addition of 3% red pepper showed the strongest inhibitory effect(27.71%).However,the star anise and myrcia promoted the formation of ?-Carboline HAAs.The inhibition effect of green pepper and cinnamon essential oil on ?-Carboline HAAs in braised meat were weak(1.5%-17%).Furthermore,the scavenging ability of HO radical was related to the inhibitory ability of spices,which provided clues for studying the inhibitory mechanism in the future.(3)Effect of spices polyphenols on the formation of ?-Carboline HAAs in a chemical modelTryptophan was an essential precursor for ?-porphyrin HAAs.Phenylalanine,glucose and creatine were not essential precursors for ?-carboline HAAs,but the addition of glucose promoted the production of Norharman and Harman.As the temperature and heating time increased,the content of HAAs in the tryptophan and tryptophan+glucose models increased,the color of the model system deepened,and the browning and the pH increased.After LC-MS analysis,the main polyphenols in ginger and pepper were curcumin,rutin,quercetin and chlorogenic acid.In the tryptophan model,at the addition of 0.05 mmol,the four polyphenols significantly inhibited the production of Norharman and Harman,and the inhibition rate was over 70%,and Quercetin has the strongest inhibitory effect.(4)Study on the mechanism of inhibiting ?-Carboline HAAs in the chemical model of spices polyphenolsTHCA was a characteristic intermediate of ?-Carboline HAAs by LC-MS analysis.The addition of 0.05 mol/L curcumin,rutin,quercetin and chlorogenic acid had different effects on THCA in the model.Curcumin had the strongest inhibitory effect on THCA(85.6%),the inhibitory effect of rutin and quercetin was slightly weaker(70.7% and77.8%,respectively),but chlorogenic acid can slightly promote THCA.4 polyphenols have weaker scavenging effect on THCA.The removal rate of adding 0.005 mol/L polyphenol is less than 10%,indicating that polyphenols do not inhibit heterocyclic amines through scavenging THCA.Compared with the control,the four polyphenols had significant inhibitory effects on carbonyl compounds,and the inhibitory rate was more than 90.3%.Aerobic participation can promote the formation of HAAs,and the production of HAAs was low under the condition of nitrogen filling.Chlorogenic acid and rutin have weak scavenging capacity for Norharman and Harman,while curcumin and quercetin can react with Norharman and Harman to significantly reduce the content of ?-carboline HAAs.