Separation And Characterization Of Antifungal Active Ingredients In Salix Babylonical L

[Objectivel We chose willow leaf as the research object to discover the chemical composition of the willow leaves extraction, separation and purification, and test its antifungal activity, getting the results that willow leaves have anti-fungal activity of monomer components; compounds were isolated structure identification; expedition willow leaf fungi antibacterial monomer components and sterilization cases; the separated monomer components each month (with leaves) were investigated.[Method] willow leaf with 8 times the amount, concentration of 50% ethanol extract by reflux extraction, concentrated under reduced pressure to recover the solvent extract made; extract The extract was washed successively with 2 times the amount of petroleum ether, ethyl acetate esters, n-butanol and extracted 3 times each; ethyl acetate layer was extracted with n-butanol layer obtained macroporous resin concentration was 10%,30%,50%,70%,90% ethanol in accordance with Elution polarity; and the polarity of each eluted fraction was measured inhibitory effect on fungi screened antifungal effective parts; effective fraction repeated column chromatography through silica gel to give a monomer component consistent with the requirements of purity; monomer physical and chemical inspection knowledge, HPLC purity analysis, H spectrum and C spectrum analysis to determine the monomer structure; willow leaves the determination of the three kinds of monomer components resulting fungal inhibition by agar diffusion method and in vitro drug based method and sterilization cases and determine the minimum inhibitory and bactericidal concentrations; each component was measured by high performance liquid chromatography content, conduct precision, reproducibility, stability and recoveries and other methodological study, the results after the passing of the separation a monomer component prepared by the standard curve, measured peak area, and calculate the content.[Results] weeping willow leaf through reflux extraction, petroleum ether, ethyl acetate, n-butanol extraction, purification by macroporous resin, and eluted fractions were each antibacterial test, get a 50%component of willow leaf fungi Inhibition; silica gel column chromatography to separate the components of 50% to obtain three compounds, after structural identification, analysis of structure of the two compounds, namely DMY and quercetin, and the rest of a structure is not identified; in vitro experiments showed that compound 1 and DMY fungi have significant inhibitory effect of compound 1 on fungi minimum inhibitory concentration of 0.125 mg · ml-1, the minimum bactericidal concentration of 0.25 mg · ml-1; DHM minimum inhibitory concentration for fungi 0.016 mg · ml-1, the minimum bactericidal concentration of 0.031 mg · ml-1. By high performance liquid components on willow leaves of precision, repeatability, stability and recoveries and other methodological study results showed that:the method is feasible and effective. Determination of the active ingredient, the results showed that:Compound 1 in May the highest level reached 4.5μg/g, compound 2 in June the highest level reached 3.35μg/g, compound 3 in September the highest level reached 15.2μg/g.And initially established method for the determination of the components of the weeping willow leaves. willow leaf is deciduous in October, the active ingredient content is low availability. The anti-fungal effective willow leaf month is June.