Isolation And Purification Of Polysaccharides Of Ganoderma Lucidum And Injection Exploit

As a precious medicinal material for thousands of years history, Ganoderma lucidum has very high medicinal value. Ganoderma lucidum fruit body was used as the raw materials to extract polysaccharides, the purification method of polysaccharides was studied, andt the injection of Ganoderma lucidum polysaccharides(GLP) was exploited in this paper. The physicochemical qualities, allergic reaction and antifatigue effect on mice were used as index to estimate the security of GLP injection. The test results provided basic dates for application of Ganoderma lucidum polysaccharides. The test results were showed as following.(1) The optimal extraction technology of soluble polysaccharides in Ganoderma lucidum was studied. On the basis of single-factor experiments, the optimal extraction technology was determined through orthogonal and response surface methodology experiments,and the optimal extraction process was to use a material/solvent ratio of 1/28 at 84℃ for 2 h, the extraction rate reached to 1.26%. In addition, the effects of ultrasonic and microwave assisted on the extraction rate of Ganoderma lucidum polysaccharides were studied, the result showed that ultrasonic and microwave could improve the extraction rate of polysaccharides notably.(2) The method of protein removal from crude GLP was researched, the optimal method was zymohydrolysis-Sevage and the test results was showed as following:papain, trypsinase,and pronase concentration were 0.8%,0.8% and 1.0% respectively, enzymolysis time was 4hours, the rate of removed proteins reached to 99.64%.(3) Deproteinizations GLP was isolated and purified by DEAE-52, and Main polysaccharides was got, it could be eluted by 0.1 and 0.5 mol/L NaCl solution in DEAE-52 column chromatography accorded to the main peak of NaCl solution concentration.(4) Main polysaccharides identified by Glucan G-150 gelatin chromatographic analysis was pure fractions,and the Deproteinization GLP have no absorption at 280 and 260 nm, wich indicated that the protein,polypeptide and nucleic acid had been removed from GLP.(5) According to the analysis of IR fingerprint characteristics absorption, the result showed that the GLP was a kind of acidic polysaccharides containing a carboxyl, and has typical polysaccharides characteristic peak.(6) Combined with hot water diffusion, Sevage’s methods, ion-exchange liquid chromatographic column and dialyze,the GLP impurities could be removed entirely, and it’s could be made into definite,high purity,according with the stipulated quality of Ganoderma lucidum polysaccharides injection.(7) The negative result of guinea pig’s allergic reaction showed that Ganoderma lucidum polysaccharides injection was safety. Intravenous injection had no allergic reaction, according with the stipulated quality of injection.(8) Ganoderma lucidum polysaccharides injection could obviously promote the antifatigue of mice, and obviously lengthen the load swimming time of mice too.The load awimming time of high-dose could be extended 85.8%. At the same time, effect of Ganoderma lucidum polysaccharides injection on ponderal growth was not notability.