Study On The Extraction Of Antioxidant Components And The Digestion And Absorption Of Phenolic Components In The Fruit Of Aroni Melanocarpa

This study is based on the Aroni Melanocarpa ftuit as the research object,optimizing the process of ultrasonic auxiliary ethanol-ammonium sulfate dual-water phase system to extract its antioxidant component and verifying its protection of oxidation stress HepG2 cells through cell testing;In vitro simulated digestion tests were carried out on the crude and purified products of Aroni Melanocarpa ftuit,and the changes of polyphenol content and antioxidant activity of Sorbus nigricans fruit were determined;Finally,the absorption law of the representative polyphenols in the fruit of Aroni Melanocarpa was studied by using the Caco-2monolayer cell model.The main findings are as follows:（1）The ultrasonic-assisted ethanol-ammonium sulfate aqueous two-phase system was used to optimize the extraction process with ABTS⁺·and·OH removal rate as the inspection index:the ratio of material to liquid was 1:21 g/m L,the mass concentration of ammonium sulfate was0.31 g/m L,the volume fraction of ethanol was 43%,the ultrasonic power was 510 W,and the ultrasonic time was 50 minutes;Under these conditions,the·OH clearance rate of the extract was（52.64±0.76）%,ABTS⁺·clearance rate was（66.08±0.84）%,and the average clearance rate was（59.36±0.80）%,which was significantly higher than that of conventional ethanol extraction and single aqueous two-phase extraction（P<0.05）;The IC₅₀ values of the extracts for ABTS⁺·,·OH and DPPH·clearance were 0.614,2.273 and 0.386 mg/m L,respectively.Through Pearson method analysis,the results showed that there was a strong correlation between polyphenols in the extract and free radical scavenging rate.（2）In order to further study the antioxidant activity of polyphenols from the fruit of Aroni Melanocarpa.The oxidative stress model was established by the action of mol/L H₂O₂ on HepG2 cells for 4 hours,and the protective effect of polyphenols from the fruit of Aroni Melanocarpa on the oxidative stress cells was investigated.The results showed that after（10-300）μg/m L polyphenol extract from the fruit of Aroni Melanocarpa can effectively improve the survival rate of cells,reduce cell damage caused by oxidative stress,and maintain normal cell morphology;Compared with the model group,low（50μg/ml）,medium（150μg/ml）,and high（300μg/ml）concentration of black fruit gland ribs of ribs with polyphenol treatment cell SOD and GSH-PX are treated with polyphenol treatment The vitality is significantly improved(（P<0.05）,the CAT vitality of the medium and high concentration group has significantly increased（P<0.05）,and the content of oxidized metabolic products MDA and protein cymbal group decreased significantly（P<0.05）,it showed that the polyphenols from the fruit of Aroni Melanocarpa could alleviate oxidative stress damage.（3）The changes of polyphenol content and antioxidant capacity of Aroni Melanocarpa fruit during digestion were investigated by simulated digestion in vitro.The results showed that the changes of polyphenol content in bold and purified products were not significant after simulated gastric digestion,but the polyphenol content in bold and purified products were seriously lost after intestinal digestion,with the loss of 19.86%and 30.48%,respectively.After simulated gastric digestion in vitro,the scavenging capacity of·OH,DPPH·and ABTS⁺·of the bold and purified substances had almost no change compared with that before digestion,while after intestinal digestion,·OH,DPPH·and ABTS⁺·had decreased to varying degrees.（4）The preliminary experiment showed that chlorogenic acid and neochlorogenic acid were the main monomer components in the polyphenol extract of Aroni Melanocarpa,,and had a high correlation with antioxidation.In order to further study the absorption law of both in the body,the Caco-2 monolayer cell model was used to simulate the transport test.The results showed that the transport amount of the two compounds was dependent on the concentration and time of the extract,and increased with the increase of concentration and time;The apparent permeability coefficients of neochlorogenic acid and chlorogenic acid in the cell model are greater than 1×10^-6 cm/s,can be well absorbed by the body,and the two substances are mainly transported passively,but the absorption effect of neochlorogenic acid in the body is better than that of chlorogenic acid.