| Object:Based on the traditional Chinese medicine compound gongyanjing were extracted by differentsolvents,to determination of antibacterial activity in vitro of various extracts of different concentration,Determining have better effect of the extract and its concentration in vitro bacteriostasis; By activity test,and ultimately determine the best solvent for the extraction of Chinese medicine and the best dosage;Determination of the effective components of traditional Chinese medicine compound Gong Yanjing, UsingL9(34) orthogonal design method to select the best extraction process of total alkaloids in traditionalChinese medicine compound gongyanjing. To lay the foundation for further development and utilizationoftraditional Chinese medicine compound Gong Yanjing and developing new dosage forms.Methods:Experiment1: The Gong Yanjing compound was extracted by petroleum ether,70%ethanol anddistilled water, by agar punch method, By comparing the size of inhibition zone to determine theantimicrobial activity of different solvent extract against Staphylococcus aureus, Escherichia coli,Salmonella, Streptococcus. Then the Staphylococcus aureus, Escherichia coli as the experimental bacteria,Determination the minimum inhibitory concentration of compound alcohol extract and water extract, Andto investigate the effects of culture medium pH, sugar, temperature and ultraviolet to bacteriostatic activityof alcohol extract and water extract. Experiment2: Were given the mice saline, dexamethasone, and high,medium and low doses of petroleum ether extract, ethanol extract, water extract, Ig four days later, with thexylene-induced inflammation, measured mouse ear swelling rate. And injection of0.7%acetic acid inducedinflammation, measured OD value of peritoneal fluid at a wavelength of590nm. Experiment3: To establishear edema model in mouse induced by xylene, egg white induced paw edema model in mouse and tamponsgranuloma model in mouse. By measuring the mouse ear swelling rate, toe swelling rate and granulationtissue weight caused by tampons to determine the anti-inflammatory effect of ethanol extract. Experiment4:The experimen by exploring ethanol concentration,ultrasonic extraction times, ultrasonic extraction timeand ratio of material to liquid, the single factor effect on the extraction rate of experimental extraction,stachydrine hydrochloride as standard, Using L9(34) orthogonal design method to select the best extractionprocess of total alkaloids in traditional Chinese medicine compound.Results:Experiment1: The alcohol extract, water extract had a good antibacterial effects. The compoundalcohol extract extract for Staphylococcus aureus minimum inhibitory concentration is0.00625g/mL, theminimum inhibitory concentration for E. coli and B. subtilis is0.125g/mL the minimum inhibitoryconcentration of water extract for S. Aureus and Escherichia coli is0.125g/mL. Temperature andUltraviolet rays treatment on compound ethanol extract, water extract did not affect on the inhibitory effect;With the increase of sugar content in culture medium, the antibacterial activity of two extracts showedincreasing trend, Two solvent extracts antibacterial activity weakened with medium pH values increased.Experiment2: Ethanol extract of ear swelling each dose group was significantly lower than the saline groupand petroleum ether group (P<0.01), and medium dose extrac of ethanol effects was the best; Ethanolextract of each group and dexamethasone group peritoneal capillary permeability were significantly smallerthan groups outside the dexamethasone group (P<0.01), And ethanol dose group was significantly lowerthan in the dexamethasone group (P<0.05). Showed ethanol was the best extraction solvent extractas activeingredient of the compound. Experiment3: Medium dose ethanol extract could significantly inhibit thexylene-induced ear swelling rate and significantly lower than the ethanol extract of high and low dosegroup (P<0.05), and compared with the dexamethasone group had no significant difference (P>0.05);Ethanol extract could significantly reduce the rate of egg white induced paw edema in mice, and the dose ofethanol extract group compared with the saline group were significantly different (P<0.01) andsignificantly lower than the high and low doses of ethanol extract group (P<0.05), compared with thedexamethasone group had no significant difference (P>0.05); Ethanol extract could inhibit the proliferation of granulation tissue in mice tampons, compared with the saline group were significantly different (P<0.01),significantly lower than the high and low doses of ethanol extract group (P<0.05) compared with thedexamethasone group had no significant difference (P>0.05). Experiment4: Volume fraction of70%ethanol, ultrasonic extraction time was25min, ultrasonic extraction three times, solid-liquid ratio of1:10,is the traditional Chinese medicine Gongyanjing best extraction technology of total alkaloids. And In all thefactors of extraction of alkaloids in traditional Chinese medicine compound gongyanjing, Influencefollowed by solid-liquid ratio, solvent concentration, ultrasonic extraction times, extraction time.Conclusion:With70%ethanol as solvent extract has better antibacterial effect in vitro and in vivoanti-inflammatory effect, Volume fraction of70%ethanol, ultrasonic extraction time was25min, ultrasonicextraction three times, solid-liquid ratio of1:10, is the traditional Chinese medicine Gongyanjing bestextraction technology. |
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