| Flavonoids is a kind of secondary metabolites present in various plants and edible fungi,and have many active effects.It has a certain regulatory effect on the human immune mechanism and cancer prevention and control system,and can achieve a series of anti-oxidation functions by eliminating a large number of free radicals.It also has a series of functions such as lowering blood fat and resisting fatigue,so it is relevant at home and abroad.The field of academic research has attracted much attention.In recent years,studies on the flavonoids of auricularia auriculatus have focused on extraction and antioxidant properties,and studies on the purification and structure of auricularia auricula flavonoids have never been reported.In this study,wild auricularia auricula with high food and medicine value was selected as the experimental material,ultrasonic extraction was used to extract the flavonoids,and the extract was separated and purified.Liquid chromatography tandem mass spectrometry(LC-MS)was used to purify the solution.The flavonoids contained in the structure and type of preliminary identification,and its anti-oxidation activity also made a related study.The results of the study are as follows:(1)Prior to the start of the experiment,the main measurement methods were evaluated and tested.The results showed that all indexes(precision,reproducibility,stability,and recovery)were qualified,demonstrating the main measurement methods used in this experiment have a certain degree of reliability.(2)Using ultrasound-assisted method combined with analysis methods of single factor and orthogonal test,we obtained the optimum extraction process of auricularia auricula flavonoids: ethanol concentration 80%,solid-liquid ratio 1:50,extraction temperature 70°C,extraction time 110 min.Under this process condition,the extraction rate of flavonoids in auricularia auricula reached 0.68%.(3)Selecting a resin suitable for purifying auricularia auricula flavonoids by comparing the static adsorption rate,static desorption rate,adsorption amount and desorption amount between 60-100 mesh and 100-200 mesh polyamide resins.The results showed that the 100-200 mesh polyamide resin had good desorption effect on the auricularia auricula flavonoids.(4)In the 100-200 mesh polyamide resin purification experiment,the adsorption and desorption experiments were carried out respectively,and the four sample indexes were selected: sample loading pH,sample volume and resin ratio,ethanol concentration and ethanol elution volume.analysis.The results showed that when the sample solution pH 6-8,the sample volume and resin ratio of 12:1,ethanol elution concentration of 90% and the elution amount of 8BV,the purification effect of auricularia auricula flavonoids reached optimal.(5)The purified auricularia auricula flavonoids were separated and identified by liquid chromatography tandem mass spectrometry.There were 18 compounds had been found in auricularia auricula flavonoids.It is speculated that the 12 compounds may be spironosine-6-sultanoyl ester,vitexin-2-O-rhamnoside,quercetin-3-rutinoside-7-galactoside,and new aconite Terpene flavanone,epigallocatechin-3-O-gallate,thiotin D,2-isoprenyl-6a peapea,dehydrocorbetone,7-hydroxy-3,4-Methylenedioxy-8-C-isoalkenoflavone,cyclamidine,rhamnoether-3-galactose-(1-4)-galactoside and 5-hydroxy-7-iso Alkenyl-8-C-isoalkenyl flavone.Due to the lack of comparison of standard products,the remaining six substances need further confirmation.(6)The auricularia auricula flavonoids were extracted by the use of a polar solvent such as petroleum ether,and the content of flavonoids in the four extraction phases of auricularia auricula was obtained: the ethyl acetate phase contained the highest amount of total flavonoids,which was 12.94%,the content of alcohol phase was the second one,and the content was 9.76%,the total flavonoids content in the chloroform phase and petroleum ether phase were less,the content was 7.33% and 7.20%,respectively.On this basis,the antioxidant effect was measured.The results showed that the four extractive phases of the auricularia auricula flavonoids had a certain ability of scavenging free radicals and anti-oxidation,and its effect was positively correlated with the concentration of the sample solution. |
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