Extraction Of Bacteriostatic Compositions From Abutilon And Study On Its Antimicrobial Activity

Plants are the treasure trove of enriched biologically active substances in nature,and the active substances with antimicrobial property are widely distributed in it.People have found that there have a lot of active ingredients with antimicrobial activity in plants,which mainly including Flavonoids,Saponins,Organic acids,Steroid,Naphtha,Anthraquinones,Terpenoids,Alkaloids,etc.Abutilon(Abutilon theophrasti Medic.),which belongs to malvaceae plants,having more than 150 species all over the world,and can effectively used in the treatment of Gonorrhea,Rheumatism,Diarrhea and other diseases.Abutilon has been widely used in folk,in order to fully develop and utilize the rich plant resources,there have numerous reports about ecological mechanism both at home and abroad,however,reseaches focused on the antimicrobial properties,antibacterial mechanism of Abutilon are rarely reported.In this paper,we extracted and filtrated the non-volatile components and volatile components from the Abutilon,and investigated the bacteriostasis effects of active ingredients in vitro.In this paper,the non-volatile constituents and volatile oils were extracted respectively from the abutilon stem,and evaluates the antibacterial activity of extracts.We identified the main ingredients in the volatile oil by GC-MS expenriment;and separated the the volatile component with bacteriostatic activity by using column chromatography,and identified the structure of some materials.(1)Extracting the volatile oil from the stems and leaves of Abutilon through microwave assisted distillation extraction device,and adopted the response surface method to optimize the extraction processes.We obtained the best extraction technology conditions,which the material liquid ratio is 1:12,microwave power is 385 W,extracting time is 10 min.In this condition,the extraction rate of volatile oil are 0.183%.(2)Analyzing the chemical compositions of volatile oil which extracted from Abutilon by GC-MS analysis,the results showed that it contains alcohol?ester?ketone and free carboxylic acid,phenolic components,and three ingredients have high levels of content,which are plant(Phytol):(E)-3,7,11,15-methyl-2-16 carbene-1-alcohol content as high as 29.20%;Secondly,propionic acid,2-methyl-3-hydroxy-2,4,4-three methyl amyl acetate(Propanoicacid,2-methyl-3-hydroxy-2,4,4-trimethylpentyl ester),content of 12.81%;9,12,15-18 carbon triene acid(Z,Z,Z)-9,12,15-Octadecatrienoic acid,content is 11.22%.(3)Evaluating the bacteriostatic activity of extracts which extracted from the stems and leaves of Abutilon with n-hexane,ethyl acetate and dichloromethane solvent respectively,the bacillus subtilis is selected as the model bacterial,we found that the best bacteriostatic effect is from dichloromethane which has the strongest polarity,and the diameter of bacteriostatic ring reached 12.17 mm.(4)Separating the extracts using dichloromethane as the extract solvent by combining silica gel column chromatography and the preparation liquid chromatography,and detached three ingredients with strong bacteriostatic activity,which are belladonna pavilion,caffeic acid methyl ester and ferulic acid methyl ester.(5)We selected the The sample were determined by ultraviolet absorption spectrum,mass spectrometry,nuclear magnetic resonance(NMR)spectroscopy.It inferred the structure of the antibacterial compounds for methyl caffeic acid,ferulic acid methyl ester,belladonna pavilion according to the graph.The sample and standard were compared and analysised by HPLC and mass spectrometry,which verify the structure of the sample.And it determined the MIC of the standard of escherichia coli,bacillus subtilis and agrobacterium,caffeic acid methyl ester MIC of three kinds of bacteria were 0.31 mg/mL,0.16 mg/mL,0.156 mg/mL,ferulic acid methyl ester MIC of three kinds of bacteria were 0.31 mg/mL,0.16 mg/mL,0.156 mg/mL,belladonna MIC pavilion for three kinds of bacteria were 0.31 mg/mL,0.16 mg/mL,0.156 mg/mL.