| Spinacia oleracea is a typical green leaf vegetable of the spinach genus in the amaranth family.Its leaves are hypertrophic and have a lot of leaf flesh.In addition to rich nutrients.Spinacia oleracea is also rich in flavonoids,which is a good raw material for flavonoids.There is some research on flavonoid extraction from spinach,and to extract further purification,composition analysis and properties and study also not reported,in order to develop and utilize the resources,this paper adopts the cellulose enzyme extraction,ultrasonic extraction,cellulose enzyme combined ultrasonic extraction spinach three methods to extract the flavones,optimize extraction method,and the extracts of purification,the antioxidant activity and purification compositions containing ingredients are analyzed,the results are as follows,1.Extraction of flavonoids from spinach.Through the organic solvent screening experiment of ethanol,methanol and acetone,the extraction rate of methanol was determined to be up to 10.55%.Therefore,methanol was selected as the organic solvent for the whole experiment.Ultrasonic extraction method by single factor and response surface experiments the optimum conditions for the extraction of extraction temperature 40℃,time of 45 min,than the rising of material liquid,extraction yield of 13.05 mg/g.Cellulase extraction by single factor and orthogonal test to determine the best conditions for the adding amount of 1.5 U/g,extraction temperature 40℃,the material liquid than 1:50,enzymolysis pH5.0,extraction rate is 14.10%.Cellulose enzyme combined ultrasonic extraction: the best condition for extracting temperature 60℃,extracting time 45 min,methanol concentration 70%,enzymolysis pH5.0,adding amount of 1.2 U/g,the material liquid than 1:30,extraction yield of 15.56%.Three methods were compared to determine the optimal extraction method for cellulase combined with ultrasonic extraction.2.Purification of flavonoids from spinach.The adsorption and analytical properties of AB-8,D101 and ADS-17 macroporous resins on total flavonoids in spinach were compared by adsorption and analytical experiments,and AB-8 macroporous resins were optimized for the purification experiment.The optimal purification conditions were as follows: extraction concentration of 3mg/mL,extraction concentration of pH3.0,concentration of analytical solution of 70%,and concentration of analytical solution of pH5.0.3.Analysis of flavonoids in spinach.The pure obtained spinach flavones and the mixed standard solution containing seven flavonoids including gallic acid,chlorogenic acid,catechin,rutin,apigenin,kaempferol and quercetin were separated on the chromatographic column.According to the retention time of the mixed standard,it can be seen that the spinach flavonoids contain seven flavonoids in the mixed standard,the measured contents were: 0.83,1.37,0.64,1.46,2.12,1.25,1.87 mg/g.The standard mixture is injected at different concentrations under specific conditions,it is concluded that there is a good linear relationship between peak area and sample concentration.The results of methodology verification show that the precision of the instrument,the stability of the test sample,the repeatability of the experiment and the accuracy of the experimental method are all in good condition.4.Antioxidant activity of flavonoids from spinach.The scavenging ability of spinach flavonoids on hydroxyl radical,DPPH radical and superoxide anion radical showed a good dose relationship with the sample concentration.The order of hydroxyl radical scavenging ability was,VC> refined spinach flavonoids > crude spinach flavonoids.DPPH free radical scavenging ability order,refined spinach>VC flavonoids > crude spinach flavonoids.In the determination of superoxide anion radical scavenging rate,the scavenging effect is poor. |
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