| Objective:To study the optimal extraction process and deproteinizati on method of polysaccharides from Fomes officinalis Ames,to optimize t he extraction process,and isolate and purification process conditions.To research the antitussive and expectorant effects of the purified polysacc harides from Fomes officinalis Ames,for Fomes officinalis Ames provi de theoretical basis for in-depth research and development and utilizatio n of polysaccharide.Methods:(1) To establish a content determination me thod.Using D-anhydroglucose as a reference,determine the maximum a bsorption wavelength,determine the absorbance of the sample by phenol-sulfuric acid method,the absorbance of the sample was determined by phenol sulfuric acid method and ultraviolet spectrophotometry.(2)The con tent of polysaccharide was determined by phenol-sulfuric acid method.On the basis of single factor experiment,the total polysaccharide conten t was selected as the evaluation index.The numbers of extraction,extrac tion time,soaking time and water addition amount were taken as the m ain factors to optimize water extraction process by Box-Behnken respons e surface methodology.(3)Purification process we used anion exchange chromatography in polysaccharides from Fomes officinalis Ames;using Se phadexG-100 to column further purified.(4)Using ultraviolet pectrophoto metry and Acrylamide gel electrophoresis analysis of the purity of pol ysaccharide from Fomes officinalis Ames.(5)The pharmacodynamic reser ach of the Uniform polysaccharide from Fomes officinalis Ames.The anti tussive and expectorant experiments were carried out with the method o f ammonia water induced cough and phenol red excretion in mice.The antitussive rate and expectorant index were selected as the indexes to o bserve the antitussive and expectorant effects of the Uniform polysaccha ride from Fomes officinalis Ames.Results:(1)The linear relationship bet ween D-anhydroglucose in the range of 0.02 to 0.06 mg has a good,r=0.9993,the average recovery was 101.63%,and the RSD value was 0.34%.(2)The optimal water extraction process was to add 10 times of w ater to extract one hour for twice.The optimal alcohol precipitation pro cess was:concentration multiple is 15 times,adding ethanol to make the content of ethanol be 80% with standing 12 hours.(3)Two components were isolated and purified from Fomes officinalis ames with yields of 54.3%and 21.7%,respectively.(4)The ultraviolet pectrophotometry and polya crylamide electrophoresis showed that the separated and purified compon ents were all homogeneous components.(5)The uniform polysaccharide F OPS-A1 and FOPS-B1 from Fomes officinalis ames has antitussive and expectorant effects,and therapeutic effect on chronic bronchitis.Conclusio n:polysaccharide from Fomes officinalis ames has cough effect andexpec toranting action,and therapeutic effect on chronic bronchitis. |
