Purification Process Of Total Flavonoids From Licorice And Inhibition On α-glucosidase

Licorice is not only a traditional Chinese medicine in China,but also one of the substances with the same medicinal and food sources.It contains various bioactive components,with flavonoids being one of its main bioactive components.It has various pharmacological activities such as hypoglycemic anti-inflammatory,antibacterial,antioxidant,and anti-tumor,and has great application prospects in industries such as medicine,health care,and food.Due to the scarcity of wild licorice resources.,therefore,it is urgent to find an efficient and rapid method for extracting and purifying total flavonoids from licorice,protect the wild resources of licorice,and improve the utilization efficiency of licorice resources.In the past,people’s research on licorice mainly focused on the extraction process of flavonoids,and their quality evaluation was only based on liquiritin and liquiritigenin,making it difficult to comprehensively evaluate the quality of licorice.Therefore,the subject studied the purification process of the total flavonoids of licorice,determined the content of its symbolic compounds in the purified products,and finally studied the inhibitory effect of the total flavonoids of licorice onα-glucosidase by using the enzyme catalysis kinetic method,in order to provide a theoretical basis for further research on the hypoglycemic activity of total flavonoids in licorice,and also to provide reference for the further utilization of licorice resources.The main research findings are as follows:（1）Research on the purification process of total flavonoids from licorice.By comparing five different macroporous adsorption resins,the optimal macroporous resin was selected,and based on this,the purification process of total flavonoids from licorice was studied.The optimal purification process was obtained as follows:the optimal macroporous resin was DM301 macroporous tree,and the optimal purification process conditions were as follows:the concentration of the sample solution was 1.85 mg/m L,the volume fraction of the eluent was50%ethanol,the volume of the sample solution was 100 m L,and the volume of the eluent was160 m L.Under these conditions,the purity of total flavonoids in the purified product obtained through macroporous resin purification reached 45.20±1.06%,and the purity of the purified total flavonoids was 4.05 times higher than before purification.（2）Establishment of a high-performance liquid chromatography method for the simultaneous determination of four flavonoids in purified total flavonoids of licorice.Using high-performance liquid chromatography,the content of four landmark compounds（liquiritin,liquiritigenin,isoliquiritigenin,and licochalcone A）in the purified product of licorice flavonoids was determined by gradient elution,based on the investigation of fluidity system,flow rate,and column temperature conditions.The methodological investigation results showed that liquiritin,liquiritigenin,isoliquiritigenin,and licochalcone A had good linear correlation within the determination range,with linear correlation coefficients above 0.9993and recovery rates above 96%.The results indicate that this method is simple,fast,precise,and stable,and can be used to evaluate the quality of licorice medicinal materials.The established method was used to determine the content of purified samples of total flavonoids in licorice,and the average content of four compounds was 58.40mg,72.46 mg,15.98 mg,and 10.62 mg,respectively.（3）Enzyme catalysis kinetic method was used to study the inhibitory effect of total flavonoids of licorice onα-glucosidase.Using purified total flavonoids from licorice as raw material and acarbose as positive control,4-nitrophenyl-α-D-glucopyranoside（PNPG）was used as the substrate to study its enzymatic kinetics.The results showed that the purified licorice flavonoids had a certain inhibitory effect onα-glucosidase,and the inhibitory rate gradually increased with the increase of the concentration of the purified substance.The inhibitory rate of purified total flavonoids from licorice onα-glucosidase is 76.43±1.43%,and the IC₅₀was 0.040 mg/m L.The inhibitory type of licorice flavonoid purification onα-glucosidase belongs to reversible anticompetitive inhibition.This will provide a basis for the medicinal development ofα-glucosidase inhibitors of purified total flavonoids from licorice.