Study On The Extraction Technology Of Total Phenolic Acid Of Cypress Fruit And Its Anti-wound Infection

Objective:By taking the immature fruits of terminalia chebula of Tibetan medicine as the research object,the paper inspects the extraction process of total phenolic acids of the immature fruits of terminalia chebula through the response surface method;macroporous resin and silica gel column chromatography and other separation methods are used to isolate active compounds;gallic acid and ellagic acid are respectively taken as reference substances,and conducts standard research on the quality of immature fruits of terminalia chebula by chromatography;meanwhile,the antibacterial activity researches on the total phenolic acid of the immature fruits of terminalia chebula and the isolated compounds are conducted.A preliminary discussion on its mechanism is carried out so as to lay theoretical basis for the development of the immature fruits of terminalia chebula of Tibetan medicine.Methods:1.The comprehensive score of the total phenolic acids is taken as an index,the extraction process of total phenolic acids is inspected,and the response surface method(RSM)is adopted to predict the optimal extraction process.2.Macroporous resin and silica gel column chromatography and other methods are used to isolate active compounds;the physicochemical properties and spectroscopy method are used;the chemical structures of the three compounds have been identified.3.Gallic acid and ellagic acid are respectively taken as reference substances,the high performance liquid chromatography(HPLC)is used to determine the content of gallic acid and ellagic acid in the immature fruits of terminalia chebula,thus to control the quality of the immature fruits of terminalia chebula.4.The clinically isolated gram positive bacteria(staphylococcus aureus),gram negative bacteria(gas enterobacter,escherichia coli,klebsiella pneumoniae,stenotrophomonas maltophilia,pseudomonas aeruginosa,burkholderia cepacia,and enterococcus faecalis)are taken as the research objects to conduct in vitro antibacterial experiments on the total phenolic acids of the immature fruits of terminalia chebula and the isolated monomer compounds.The electron microscopy(sem)and crystal violet staining method are used to preliminarily discuss antibacterial mechanism.Results:1.The optimal extraction process was reflux extraction3.39h,the solvent was 24.67 times,and the solvent concentration was 74.66%.Combined with the actual situation,the extraction conditions were changed to be reflux extraction 3h,the solvent dosage was 25 times,the solvent concentration was 75%,the comprehensive score was 73.71.2.5 compounds were isolated and3 of them were identified;they were respectively gallic acid,ellagic acid and arjunolic pigment.3.By HPLC determination,the content of gallic acid was 1.74%,and ellagic acid content was 0.577%.4.Arjunolic elements had more obvious antibacterial effect than the other two compounds,which has obvious inhibiting effect on pneumonia gram staphylococcus aureus and Enterococcus faecalis.At the same time,with the increase of the concentration of the compounds,the number of the biofilm decreased;the dye experiment on biofilm fluorescence showed,arjunolic elements can not only decrease the number of live bacteria in the bio film and the number of dead bacteria had also been reduced.Conclusions:1.Compared with the traditional process,the use of response surface method to carry on the study of the extraction process of total phenolic acids of the immature fruits of terminalia chebula has more accurate results and higher accurancy.2.The high performance liquid chromatography method is adopted,gallic acid and ellagic ellagic acid are taken as reference substances to determine the content of Terminalia chebula herbs gallic acid and ellagic spend acid,which can be more systematically and accurately control the quality of the immature fruits of terminalia chebula.3.The antibacterial mechanism of arjunolic element may be related with the destruction of the bacterial cell membrane,and it has obvious inhibitory effect on the formation of bacterial biofilm.