Inhibitory Effect And Mechanism Of Sonchus Oleraceus Linn Extract On The Formation Of Carotoxic Heterocyclic Aromatic Amines

With the improvement of people’s living standards,the issue of food safety has become more and more important,and it has become a hot issue in the current society.Therefore,as a carcinogen closely related to human daily diet,heterocyclic amines have received widespread attention from researchers,and how to effectively inhibit the production of heterocyclic amines has also become a hot research issue.As a traditional Chinese medicinal and edible plant,Sonchus oleraceus Linn has been proved to have liver-protecting,diuretic and antitumor effects,but few studies have investigated the effect of SOL on heterocyclic amines.Therefore,this project takes SOL ethanol extract as the research object,by adding it to pork mill at high temperature,roasting and pulverizing the sample,and using solid phase extraction(SPE)method to extract,enrich and purify heterocycles amines in the sample.using HPLC to quantitatively detect six heterocyclic amines(Ph IP,Me IQx,IQ,4,8-Di Me IQx,harman,Norharman),and observe the inhibitory effect of SOL extract on heterocyclic amines;reuse Creatinine-phenylalanine(1: 1,v/v)chemical model system to study the inhibition mechanism and action pathway of Ph IP by SOL extract;finally,Caco-2 induced by Ph IP,Me IQx,IQ,4,8-Di Me IQx Cells were used as models to investigate the protective effect of SOL extract on heterocyclic amine-induced DNA damage.Themain results of this paper are as follows:Firstly,the infrared spectrum and high performance liquid chromatography were used to analyze the active ingredients in the ethanol extract of SOL,and then the DPPH,ABTs,FRAP in vitro antioxidant experiments were used to determine the free radical scavenging ability of SOL extract under different temperature conditions,and the total The content of total phenol and total flavonoids,and the effect of temperature on the antioxidant capacity,total phenol and total flavonoids of SOL extract.The results showed that the samples contained polyphenolic and flavonoid groups,and the peak shape was more obvious.Using high-performance liquid chromatography to identify them,the results showed that the main substances were caffeic acid and chlorogenic acid.The increase of the antioxidant capacity of SOL extract gradually increased the total phenol and total flavones content.However,compared with the untreated control group,it was found that the total flavones and total phenols were significantly losser at 60°C heat treatment.,antioxidant capacity is also reduced.Subsequently,solid phase extraction-high performance liquid chromatography was used to detect the heterocyclic amines formed in the food system(pork)as the target to determine the inhibitory effect of SOL extract on heterocyclic amines.The results showed that the total amount of HAAs detected in the samples added with SOL extract was increasedafter adding SOL extract to roast pork under different conditions.In the normal group(200 ° C 45min)samples,the best inhibitory effect on total HAAs production was the sample with the addition level of SOL extract at 3%;The best inhibitory effect was the sample with 1.5% addition of SOL extract.And the cooking temperature and cooking time are significantly related to the formation of heterocyclic amines in the food system and the physical and chemical properties of pork samples.Lowering the cooking temperature and shortening the cooking time can effectively reduce the production of heterocyclic amines and reduce cooking losses.After clarifying the inhibitory effect of SOL extract on heterocyclic amines,the creatinine-phenylalanine(1: 1,v / v)was used as a chemical simulation system to study its inhibitory mechanism on Ph IP and to investigate seven polyphenol flavones Effects of Substances on Ph IP Formation.It was found that with the increase of the amount of SOL extract,the creatinine content gradually increased,and the phenylacetaldehyde and Ph IP content gradually decreased.Investigate its inhibitory pathway by the addition reaction of polyphenol and flavonoids in SOL extract with phenylacetaldehyde,which consumes phenylacetaldehyde in the model system,and then reduces the reaction between creatinine and phenylacetaldehyde,thereby inhibiting Ph IP form.The inhibitory effects of seven polyphenol flavonoids on Ph IP were:quercetin> kaempferol> apigenin> genistein> chlorogenic acid> caffeic acid> p-coumaric acid.Moreover,the correlation between the free radical scavenging ability of polyphenols and its inhibitory effect on Ph IP was studied,and it was found that the inhibitory effect of polyphenols on heterocyclic amine Ph IP had no significant correlation with its free radical scavenging ability(r=-0.087,p=0.517>0.05).Finally,Caco-2 cells with heterocyclic amine-induced DNA damage were used for cell experiments to analyze the protective effect of SOL extract on heterocyclic amine-induced DNA damage.It was found that DNA strand breaks induced by IQ,Me IQx,and Ph IP showed effects at SOL concentrations of 0.1-100 μg/m L,and DNA strand breaks induced by 4,8-Di Me IQx were only at lower concentrations of SOL concentration(0.1-1 μg/m L)has a protective effect,but at higher concentrations(10-100 μg/m L),no significant protective effect was observed.Moreover,the apoptosis rate of the sample group treated with SE was significantly reduced at all concentrations(p <0.05),showing that the SOL extract had a good effect on inhibiting heterocyclic amine-induced apoptosis at each concentration.