| Objective：Find the quality control indicatro and establish the qualitystandard of Lobaria kurokauae by the experimental studying on efficacy partsand the chemical composition. Provid the scientific basis for evaluatingthe quality and rationally development and application of Lobaria kurokauae.Methods：Extract Lobaria kurokauae with water and ethanol respectively.screen the efficacy parts of anti-inflammatory effects by evaluatingxylene-induced ear saelling in mice and cotton-induced granuloma in mice.screen the efficacy parts of the in vitro antibacterial activity by doubledilution of the drug in the test tube and paper disk method. screen theefficacy parts of in vitro antioxidant activity by the test of scavengingDPPH and reducing Fe3+.Use mice to study the maximum tolerated dose（MTD）of different parts from Lobaria kurokauae.Determine the ethanol extract of Lobaria kurokauae as the main efficacyparts on the basis of activity screening.Extract with80%ethanol,partitioned extract with petroleum ether, ethyl acetate and n-butanol.Separate and purify the chemical composition by silica gel columnchromatography for every extractant.Consult Chinese Pharmacopoeia (2010Edition) as the mainstay. Study theidentification items by pharmacognosy and thin layer chromatography(TLC)identification.study the inspection items by determin the content ofmoisture, ash and harmful elements.Study the content of extractum by ethanolhot extcacting. Study the method of determine the content of the maincomponent retigeric acid A in Lobaria kurokauae by HPLC.Results：Compared with the negative control group, Ethanol extract of Lobaria kurokauae at high-dose have significantly inhibition forxylene-induced ear saelling（P＜0.05）, inhibition ratio was45.38%, Ethanolextract of Lobaria kurokauae.at low-dose have very significantly inhibitionfor cotton-induced granuloma （P＜0.01）, inhibition ratio was26.28%. Waterextract and ethanol extract of Lobaria kurokauae were both sensitive forEscherichia coli and Candida albicans. the bacteriostasis effects of ethanolextract was better than water extract,both of them were inferior onsterilization. The effect of ethanol extract on scavenging DPPH and reducingFe3+were both better than water extract. The MTD of water extract for micewas148g crude drug/kg, equivalent to592times for the amount of person-days.The MTD of ethanol extract for mice was196g crude drug/kg, equivalent to784times for the amount of person-days.7compounds are separated by silica gel column chromatography, use1H-NMR,13C-NMR and MS to identified structures.they are palmitinic acid,Stictan-3β,22α-diol, oleanolic acid, ursolic acid, aurantiamide acetate,retigeric acid A and retigeric acid B. Oleanolic acid, ursolic acid andaurantiamide acetate are the first time separated from Lobaria kurokauae.Determine the TLC condition for identification items, retigeric acidA and retigeric acid B are comparison, developing solvent is toluene-ethylacetate-formic acid. Determine inspection items, moisture content is notmore than12.0%, total ash is not more than14.0%, acid insoluble ash isnot more than8.0%, lead is not more than25ppm, cadmium is not more than0.3ppm, copper is not more than12ppm, arsenic is not more than2ppm,mercury is not more than0.6ppm. Determine the content of extractum byethanol hot extcacting is not less than6.0%. Determine the content ofretigeric acid A by HPLC is not less than0.43%.Conclusion：Ethanol extract is the mian efficacy parts of Lobariakurokauae for anti-inflammatory, in vitro antibacterial and in vitroantioxidant activity. the clinical dosis is security. Further understoodthe chemical composition of Lobaria kurokauae and reveal the fundamental substance initiatory, provided the effective basis for rationallydevelopment and application. the established quality standard improved thelevel of quality control, provide a scientific basis for the protection ofthe safety and effectiveness in clinical applications. |
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