Study On Pharmacognosy Of Gloeostereum Incarnatum S.Ito Et Imai

The authenticity,validity and security of Gloeostereum incarnatum were studied systemically in this dissertation.In the origin,Gloeostereum incarnatum was a kind of fungi which belong to Basidiomycota,Hymenomycetes,Aphyllophorales,Meruliaceae,Gloeostereum.The dry fruitbody can use as medicine.By the macroscopic identification,the fresh fruitbody of natural G.incarnatum was pectic and like ear.Its diameter was 2-10(13) cm,the thickness was 0.5-1cm.There were many floss on the superstratum and the colour was yellow.The thickness of the floss was 1mm.The colour of the inverse was from ivory-white to jacinth.There were a lot of vesicle tuber distribute radially on it,the diameter was 1×1-3mm.The fracture section was pectic,and the colour was cuticolor.Both the taste and smell were weak.The differences of culture G.incarnatum were that the size of fresh fruitbody of cultural G.incarnatum were average.The diameter was 4-8cm, and the floss was 1.5-2mm.The inverse was jacinth.The dry fruitbody of natural G.incarnatum was hardness,and the diameter was 2-3(5)cm. The fracture section was uneven,and smell like mushroom.The changes were not remarkable when after soak in water and open-air drying.The differences of culture G.incarnatum were that the size of dry fruitbody,the diameter was 4-7cm,which is doubled to quintupled after they are soaked in water.They are changed into lamelliform after open-air drying.On the producing area,natural G.incarnatum was distributed at the Tibet Autonomous Region and Jilin Province mainly at the present time.Thereinto,most of them were came from Yehe Town Siping City.The distrbuting was thin in the other places of Jilin Province.The primary producing area of cultural G.incarnatum was Jilin Province.In this place,the annual output is 70-80%of gross,and the throughput of Jiaohe City and Siping City are more.On the harvest,natural G.incarnatum can be collected once annually.The best time to collest the fruitbodies was from July to September.Cultural G.incarnatum can be collected twice annually.June to July in spring,September to October in autumn.The fruitbodies should be insolation on the air.The microstructure of G.incarnatum was observed using paraffin slice method.The results were that the microstructure including hymenium,mesosphere and surface layer.The hyphae made the surface layer.The hyphae of mesphere had clamp-connection.There were a lot of interspaces as bladder in the mesosphere of fruitbody of G.incarnatum.These interspaces was the reason that there were many vesiculate protuberances on the hymenium.The number of basidiospores of natural G.incarnatum were more than culture G.incarnatum.On the chemistry,the dry fruitbodies of G.incarnatum were extracted with MeOH 3 times at 50℃.The extract was concentrated after combination,then save the liquor and put into the silica-gel equimultiple.Evaporate this liquor in water.Circumfluence concatenation extraction 3 times with petroleum ether,chloroform,EtOAc,acetone and MeOH,and concentrate the extract leave each other.Then the extracts were departed into 6 parts,including the petroleum ether fraction,the chloroform fraction,the EtOAc fraction,the acetone fraction,the methanol fraction and the water fraction.The petroleum ether fraction placed at 4℃72 hours;the sediment was purified with diethyl ether and gave compound 1.The chloroform fraction was subjected on silica-gel column with a Petroleum ether:EtOAc(12:0.5) elution solvent and gave 6 fractions. The D-3 was subjected on silica-gel column with a Petroleum ether:EtOAc(7:0.5) elution solvent again,then gave 5 fractions.Compound 2 was purified from D-2.The acetone fraction was concentrated and deposited for 72 hours;the sediment was purified with acetone,MeOH and then gave compound 3.Compound 1-3 were identified as ergosterol, 11—α—penteny—2,9,16—octadeca-carbon triene and Mannitol using spectroscopic methods, inclding HPLC,NMR,MS and so on.Thereinto,11—α—penteny—2,9,16—octadeca-carbon triene was a new compound.By the physical and chemical identification,the types of chemical compounds in G.incarnatum were flavonoids,cardiac glycosides.alkaloids,polysaccharides,volatile oil, triterpenes,amino acids,polypeptides,and proteins.In the phenomenon of microsublimation, there were yellow crystal like frostwork.The component of crystals was volatile oil.By the content test,it was contain proteins,fattiness.glucide and many microelements of G.incarnatum.The the pesticide residue of HCH and DDT was up to snuff which belong to natural and cultural G.incarnatum,were analyzed by GC.But the contents of Pb in natural G. incarnatum came from Yehe Town Siping City and As in cultural G.incarnatum came from Jiaohe City were excess.In the pharmacological,the bacteria growth inhibition assay of the petroleum ether fraction, the chloroform fraction,the EtOAc fraction,the acetone fraction,the methanol fraction,the water fraction and compound 2 were examined.The results indicated that each fraction had antibacterial activity to Escherichia coli ATCC25 923 and Escherichia coli E^ΠR_ï¿¡.The average inhibition ratio of MeOH fraction was highest.It compared with comparison group in a concentration-dependent manner.Except the chloroform and acetone fraction,the other groups cannot inhibit Staphylococcus aureus O₄₁ ermc.The inhibition rate of the acetone fraction(84 mg/mL) was 73.17%but not in a concentration-dependent manner.Both the petroleum ether fraction and the EtOAc fraction(50 mg/mL,25 mg/mL) can inhibit Escherichia coli E^R_cat,and the inhibition rates are higher than the gentamicin group.The inhibition ratio of the petroleum ether fraction(30 mg/mL) was highest.160.21%.It was remarkable compared with the comparison group(P＜0.05).The inhibition rate of the chloroform fraction was 95.61%at the concentration of 50 mg/mL to Escherichia coli E^R_cat.The chloroform fraction can inhibit Staphylococcus aureus S₁₉ NMA₁ in a concentration-dependent manner,and the inhibition ratio was 34.03%at the concentration of 50 mg/mL,and was remarkable compared with the comparison group(P＜0.01).The inhibition ratio of the EtOAc fraction(50 mg/mL) to Staphylococcus aureus S₁₉ NMA₁ was 39.36%and in a concentration-dependent manner.The inhibition ratio of the acetone fraction(84 mg/mL) was 49.87%in the effect to Escherichia coli ATCC25 923,and was remarkable compared with the comparison group(P＜0.01).The fraction can inhibit Escherichia coli E^R_cat,and the highest inhibition ratio was 80.10%at the centrations of 42 mg/mL and 21 mg/mL.It was remarkable compared with the comparison group(P＜0.05). All the concentrations of the MeOH fraction can inhibit Escherichia coli E^R_cat obviously.The highest inhibition ratio of them was 74.94%,but not in a concentration-dependent manner. Compound 2 put up obvious inhibition effect on Escherichia coli ATCC25 923,Staphylococcus aureus ATCC25 923 and Escherichia coli E^ΠR_ï¿¡.Thereinto,the effect on Escherichia coli E^ΠR_ï¿¡better and better with the concentration lower and lower.The inhibition ratio of Compound 2(0.011 mg/mL) was 48.78%in the effect to Staphylococcus aureus O₄₁ ermc.In conclusion. the acetone fraction,the methanol fraction,the water fraction and the chloroform fraction of G. incarnatum had obvious antibacterial activity,Compound 2 also.The water fraction of G.incarnatum has the effect on mice ears plumping induced by xylene.The inhibition rate was 15.86%at a dose of 5 g/kg,but has not remarkable difference with the control group.The authenticity,validity and security of G.incarnatum were studied systemically in this dissertation the first time,and many fractions were tested with antibacterial activity for the first time.Three compounds,including ergosterol,11—α—penteny—2,9,16—octadeca-carbon triene and Mannitol were isolated from G.incarnatum.Ergosterol and Mannitol were reported in this mushroom for the first time.11—α—penteny—2,9,16—octadeca-carbon triene was a new compound.It was not only the antibacterial activity component,but also the proper component can be used to identify G.incarnatum.The results provide scientific basis in order to establish quality standard on G.incarnatum.