Study On Anti-alzheimer’s Disease Phenolic Acids In Raspberry Based On Molecular Imprinting And Technology And Molecular Docking

Palm leaf raspberry(Rubus chingii Hu)is a Chinese medicinal material that can be used for medicine and food.It has high economic value and has been included in the Chinese Pharmacopoeia.The results of previous studies have shown that the ethyl acetate part of raspberry has anti-Alzheimer’s effect,and the flavonoids(quercetin and kaempferol)can significantly improve the learning and memory ability of naturally aging rats and dementia rats with kidney-yang deficiency.However,it is still poorly understood whether there are active compounds in phenolic acids in the ethyl acetate site.Because the traditional separation method is cumbersome to operate,consumes a lot of solvents,and is difficult to separate,it is of great significance to develop an effective separation and enrichment method to screen the phenolic acid active ingredients in raspberries.In this study,ellagic acid was used as a template to prepare surface-imprinted polymers to separate the phenolic acid components,and the activity was screened by molecular docking technology,and the content was determined by the one-test-multiple-evaluation method.1.Preparation and characterization of ellagic acid molecularly imprinted polymer:Using ellagic acid as a template molecule,the molecularly imprinted polymer was synthesized by precipitation polymerization and silica gel surface polymerization,respectively.The composition and morphology were characterized by infrared spectrometer and electron microscope.The static adsorption test and the dynamic adsorption test show that the adsorption performance of the molecularly imprinted polymer on the surface of silica gel is higher than that of the precipitated molecularly imprinted polymer.Therefore,it is used as a solid phase extractant for the separation and enrichment of ellagic acid and the like in Raspberries.2.Directional enrichment of phenolic acids in raspberry: using molecular imprintingsolid phase extraction technology,using molecularly imprinted polymer on the surface of ellagic acid silica gel to separate the activity of phenolic acids in ethyl acetate extract of Raspberry.At the same time,combined with liquid-mass spectrometry to analyze the structure of the enriched component and separate and purify the obtained compound,the structure of the obtained compound was identified by nuclear magnetic resonance and mass spectrometry.There are six components: Gallic acid,Lindenin and Kaempferol-3-O-rutinoside,Ellagic acid,Vanillic acid,and Ferulic acid.3.Screening of phenolic acid active ingredients: search for Alzheimer’s diseaserelated receptor proteins through literature and PDB database,and isolate and purify 6compounds from raspberry ethyl acetate extract based on molecular imprinting technology and anti-Alzheimer.The molecular docking of the Haimer disease protein is carried out,and the active compound is initially predicted.For screening compounds whose docking energy Lib Dock Score is greater than or equal to 0 and can interact with5 or more target proteins,they are considered as potential active compounds.4.One test and multiple evaluation method to determine the content of phenolic acid active compounds in raspberries: the content of ellagic acid in raspberries is relatively high,so this study used cheap and easily available ellagic acid as the internal standard,and adopted high-performance liquid phase.Chromatography method was established to determine the contents of gallic acid,lindenin and kaempferol-3-O-,ellagic acid and ferulic acid in Raspberries at the same time,and compared with the measurement results of external standard method.In addition to verify the durability and system applicability of the QAMS method,the applicability and accuracy of the one-test-multi-evaluation method in the multi-index quality control of raspberry medicinal materials are explored.In this study,the active groups of phenolic acids in ethyl acetate extract of raspberry were concentrated and separated by molecular imprinted polymer on the surface of synthetic ellagic acid silica gel,and six compounds were obtained,including basilitin,kaempferol 3-O-rutoside,gallic acid,ellagic acid,vanillic acid and ferulic acid.Anti AD activity was predicted by molecular docking technique.A multi-evaluation HPLC method was established to determine their contents simultaneously,which provided a theoretical and technical basis for the further development and utilization of Raspberry based on the anti-Alzheimer’s effect and the research of new Alzheimer’s drugs.