Militaris Solid Cultivation And Waste Deep Processing

The research subject had been done in depthly and systematically on extraction solid medium cultivation of Cordyceps militaris ; purification technique, detection method, construction identification of polysaccharide and cordycepin in the leftover bits, as well as the manufacture process study for comprehensive extraction, achieved satisfactory results.1.To achieving quality and high yield Cordyceps Militaris from solid medium cultivation, the excellent strain selecting, scientific compound of culture medium, careful cultivation and other segments are most important aspects.2.From the cultivation leftover of Cordyceps Militaris , hydrothermal refluxing extraction method was better than ultrasonic water extraction for cordycepin polysaccharide extraction; for cordycepin extraction, ultrasonic ethanol extraction method was little better than hydrothermal refluxing extraction,ethanol thermal refluxing extraction and ultrasonic water extraction.3.Through single factor analysis about cordycepin polysaccharide and cordycepin extraction rate, and four factors and three levels orthogonal experiments analysis, the results showed the optimum technological conditions for cordycepin polysaccharide extraction: hot reflux extraction three times at 80℃with the raw material:water ratio at 1/20 , 90 minutes each time; the optimum technological conditions for cordycepin extraction: 75% ethanol ultrasonic extraction three times with the raw material-water ratio at 1/10 , 20 minutes each time.4.Analysised process conditions that effect cordycepin polysaccharide and cordycepin extraction, and got comprehensive extraction process route by comprehensive extraction amplification experiment. The result for cordycepin polysaccharide and cordycepin comprehensive extraction from the cultivation leftover of Cordyceps Militaris: 20 times content water , extraction three times at 80℃with the raw material:water ratio at 1/20 and 2 hours each time. cordycepin polysaccharide and cordycepin Isolated from extracting solution by alcohol sedimentation, using clarifier technique to deproteinization, cellulose chromatography to purify cordycepin polysaccharide with high content; added 1% activated carbon to the alcohol precipitation supernatant ,water bath at 60℃to decoloration 30min, got cordycepin extracting solution. Anion exchange resin chromatography to achieve purification of cordycepin crude and high-quality product. Determined by HPLC and phenol-sulfuric acid method, Cordycepin polysaccharide yield of crude after deproteinization could yield 5.28% and purification higher than 50%, through anion exchange resin could yield 0.38% and purification 97%.5.Explored the cordycepin and cordycepin polysaccharide qualitative and quantitative analysis with thin layer chromatography, spectrophotometry, HPLC, infrared and other method. Cordycepin and cordycepin polysaccharide content difference had been determined by HPLC and phenol-sulfuric acid method respectively. Comparing cordycepin purification with standard used infrared method, could decided it was cordycepin.