The Basic Research Of Antimicrobial Active Substances In The Sophora Flavescens Seeds

Studies in developing countries demonstrate that high rate of antibiotics use, unreasonable use,widespread se1 fmedication of antibiotics,patients and community residents lack knowledge of properuse of antibiotics. Irrationality of clinical use of antimicrobial agents also exists in China,for example application without indication,overlong duration,overdosing or inappropriate combination of drugs,etc.Antimicrobial agents continue to account for a significant portion of hospital pharmaceutical expenditures.,so their use and evaluation for infectious diseases have important economic implications. Because of increasing of drug-resistent strain and abusing of antibiotic,studies about Tranditional Chinese Medicines which have antibacterial effect have already become a hot focus in medical field.Because of the rare number and unsustainable utilization of sophora root as Chinese medicinal materials, this topic can choose the sophora seeds as the research object.Combined with the antibacterial effect, studies include the composition,the extraction, separation and purification.The main research contents and results are as follows:1. With the anti-bacterial effect as the experimental index,among petroleum ether, chloroform, ethyl acetate, anhydrous ethanol, 80% ethanol solution and among ultrasonic extraction, pressurized extraction, reflux extraction, microwave extraction,the best extraction solvent and method that exacted antibacterial active ingredients of sophora seeds were choosed. The results showed that the petroleum ether extracts, chloroform extracts, ethyl acetate extracts of sophora seeds had no bacteriostasis, and the antibacterial activity of 80% ethanol extracts was the strongest, while the others were weaker,and the order of antimicrobial effect of four methods’ extracts from strong to weak was Ultrasonic extraction >Pressure extraction > Microwave extraction > Reflux extraction.2. Under the same experimental method, the compositions of sophora seeds were preliminary compared with the ones of sophora root. The TLC and antibacterial experiments showed that sophora seeds and sophora root had significant differences in compositions,and Sophora root and seeds all showed strong antibacterial activity in the activitygram-positive bacterium.The MIC of 80% ethanol extracts of sophora seeds was in the range of 5 mg/ml- 10 mg/ml.3. 80% alcohol extracts of sophora seeds and root were extracted as petroleum ether extracts, ethyl acetate extracts and n-butyl alcohol extracts. A variety of methods were used to analyse the compositions and activity of three layer materials. Chemical identified methods showed that the ingredients in the sophora root and seeds had obvious differences.The TLC showed that under the Pressurized extraction and Ultrasonic extraction, the extracts of sophora seeds had almost the same chemical compositions, but compared with the ones of sophora root varies greatly. Alcohol extracts of Sophora seeds didn’t contain weak polar alkaloids, and strong polar alkaloids had also obvious differences between sophora root and seeds. The HPLC showed that the ethyl acetate layer of alcohol extracts of sophora seeds contained mostly weak polar ingredientsis, but sophora root contained compositions with wider polar range. The antibacterial experiments showed that the ultrasonic extraction and pressure extraction had not influence on the extraction of bacteriostatic active ingredients in sophora seeds, and the extracted ethyl acetate layer had certain bacteriostatic effect.4. The ethyl acetate layer of 80% alcohol extracts of sophora seeds by silica gel column chromatography, gel column chromatography and preparative liquid chromatograph was separated and purified, and five monomeric compounds were acquired, using uv, mass spectrometry, nuclear magnetic methods,that were identified as:(1) β-sitosterol;(2)Biochanin A;(3)Pratensein;(4)Daidzein;(5)3’,4’-dihydroxy-7-methoxyisoflavone(unknown compound); Grease-like compositions by GC-MS were detected to get 48 componental signals, and the linoleic acid was its main ingredient(32.76%). Whether the rest of the low polar components were the new compound or not need further research.To sum up, as to the sustainably utilizated medicinal herbs, this topic preliminarily research on the antibacterial active ingredients of sophora seeds, and determining the differences between the sophora root and seeds, separating and purifying sophora seeds,geting antimicrobial active monomeric compounds, so as to conclude and summarize the internal relations between antibacterial activity and structure and resist the regeneration and growth of drug-resistant bacteria to open up a new path.