Study On The Effective Constituents And Antibacterial Activities Of Dicranostigma Leptodum(Maxim.) Fedde's Alkaloid

Dicranostigma leptopodum(Maxim)Fedde,DLF,A is biennial or perennial herbage, which belongs to Dicranostigma in papaveraceae.It is widespread at Qin ling area and Wei shui valley in Gansu province.It is entire applying for medicine in the folk. It possess efficacy as follow: pyretolysis,detoxicating,deswelling,analgesia,massacring insect and so on, the main ingredients are various isoquinoline alkaloid which have wide biologic activity.Based on the principles of phytochemistry and modern chemical separation methods,the total alkaloids contents of the different grouth periods different organs of DLF were determined by the UV method in order to investigate their distribution of the total alkaloids. The best harvest time and ideal developing solvent was defined.At the same time, Separation,purification and initial identification about two kinds of DLF′s extract were made by means of physico-chemical determination,chromatographic fractionation and spectral analysis.Using the DLF′s alkaloid to determine antibacterial activities about five kinds of bacteria species,such as Escherichia coli,Staphylococcus aureus,Bacillus subtilis,Enterococcus faecalis,Proteus mirabilis.The results were as the follows:1.The alkaloids were extracted by alcohol from different organs during different growing periods.OD value were tested through the spectroscopy at 210nm.A analysis of variance had been carried out.The alkaloids content in roots and leaves during the same growing period did not have obvious difference(t=1,P>0.05).The alkaloids content in roots during different growing periods had significant difference(F=128.11,P<0.01),the alkaloids content in leaves during different growing periods were nearly same(F=-4.938,P>0.05).The alkaloids content in the whole DLF during different growing periods were significantly different(F=19.319,P<0.01).The alkaloids content is the highest at the end of March or beginning of April in the whole DLF,so the initial flowering stage is considered as the best harvest time.2. The two kinds of extract′structure which were separated from DLF through alcohol extraction were analysed by quartz chromatographic column,TLC test,microscope melting point test,element analyse,UV,IR,MS.Results indicated:Using 95% ethanol:Ice acetic acid:water: strong aqua(15:0.5:2.5:0.5) as the developing solvent for thin-layer chromatographic analysis,this method is most effective to separate.The spot is clear.3. In vitro antibacterial activities reserch were made through using selectively DLF′s alkaloids on Escherichia coli,Staphylococcus aureus,Bacillus subtilis,Enterococcus faecalis,Proteus mirabilis. MIC, rate,IC50 were measured. Then LSD multiple comparison were carried out. Result of DLF′s alkaloidsⅠindicated:the minimal inhibitory the minimal inhibitory concentration(MIC) is 0.18mg/mL, 0.14mg/mL, 0.30mg/mL, 0.18mg/mL, 0.30mg/mL; IC50 is 0.1407mg/mL, 0.1005mg/mL, 0.1407mg/mL, 0.2605mg/mL, 0.2607mg/mL; Result of DLF′s alkaloidsⅡindicated: the minimal inhibitory concentration(MIC) is 0.22mg/mL, 0.18mg/mL, 0.18mg/mL, 0.30mg/mL;IC50 is 0.1807mg/mL, 0.1407mg/mL, 0.1407mg/mL, 0.2605mg/mL.In conclusion, the researeh above enrich the data about the chemical constituent and pharmacologic actions of DLF,which provide meaningful basis for the further development and multiple use of relevant resources.