Studies On Optimism And Extraction Of Bacteriostat Of Sophorae Flavescentis Radix

Sophorae flavescentis radix;Phellodendri chinense cortex;Cnidii fructus;Dictamni cortex can be used as in vitro antibacterial materials in the field of research of natural plants,all of which can inhibit the growth of Staphylococcus aureus.The combined use of these four-formulations in antimicrobial applications has not been reported in relevant literature.Through the Uniform Design method,the experimental design was optimized to optimize the compounding ratio,the Response Surface Method was used to optimize the compounding extraction process,and finally the optimized extract was used for more extensive bacterial inhibition tests.The specific work is as follows:1.Uniform design method was used to design unconstrained formula experiments.The effective substances in the formula were extracted by refluxing method and their antibacterial activity against Staphylococcus aureus was determined.The regression equation was used to predict the optimal formulation ratio and image rendering;The results were verified by experiments.The experimental results were basically consistent with the prediction results.The optimum ratio obtained was Sophorae flavescentis radix cortex?Phellodendri chinense?Cnidii fructus?Dictamni cortex=1?1.808?1.163?0.110.The antibacterial effect of the optimal proportion formula at the dosage of 0.006g/mL of crude drug was significantly stronger than that of the single herbs,and the antibacterial rate to Staphylococcus aureus was up to 89.53%.2.Through the investigation of the key factors in reflux extraction,three factors that have more significant effects are selected from factors such as soaking time,refluxing time,bath temperature,liquid-to-feed ratio,and ethanol volume fraction:ethanol volume fraction,reflux time and bath temperature.The inhibition rate of the formulation against Staphylococcus aureus was used as the response value,and the CCD design experiment was used to optimize the reflux extraction process of the formulation.The final optimal extraction process was:ethanol volume fraction of80.5%,reflux time of 1.6 h,bath temperature of 95°C.The extraction process was verified,and the experimental results were basically consistent with the CCD mathematical model prediction results.Under the conditions of raw herb administration of 0.003 g/mL,the optimized active ingredients were extracted and the antibacterial rate reached 94.29%.3.According to the optimized ratio and extraction,the medicinal materials was extracted and conduct antibacterial test and MIC test on Staphylococcus aureus,Escherichia coli and Bacillus subtilis.The results show that under the optimized formula and extraction,the effect of the extract of the formula on the growth of the three bacteria is as follows:the inhibition of Staphylococcus aureus and Bacillus subtilis is basically the same,and in the high concentration,It started to produce inhibitory effects in the adaptation period and kept both at a very low level of growth.Growth inhibition of Escherichia coli started when entering the logarithmic phase,but it accelerated the growth of Escherichia coli and advanced the decay phase.In the determination of MIC,the inhibition result of Staphylococcus aureus was MIC₅₀=1.2 mg/mL,MIC₉₀=2.7 mg/mL,and the inhibition result of Escherichia coli was MIC₅₀=6.0 mg/mL,and the inhibition result to Bacillus subtilis was MIC₅₀=0.68mg/mL,MIC₉₀=2.9 mg/mL.