Explorations On Best Culture Conditions Of The Fruit Body Of Cordyceps Militaris And Optimization On Detection Methods Of Its Active Components

Cordyceps militaris(vuill.)Fr. and Cordyceps sinensis(Berk.)Sacc belong to the same genus, but different species. They have almost same clinical efficacy, so the former is hopeful to be applied to officinal field instead of the latter. The natural source of the latter is extremely limited, while the solid artificial ferment has succeeded in the former. Therefore, it has great research significance to explore and optimize the culture conditions on the fruit body of the former in order to make for its industrialized mass production in future. 1 Confirmation of the best culture condition for Cordyceps militaris fruit bodyUsed firstly orthogonal experiment in five factors and four levels to research the culture condition of Cordyceps militaris fruit body ferment. Testing and analyzing desiccant weights of the collected fruit body selected the optimum culture condition. How long the time being covered was the most important factor for Cordyceps militaris fruit body bourgeons.2 Optimization on current methods of isolation and detection Cordyceps' active components 2.1 Isolation cordycepin and optimization its detection method By study on different HPLC conditions, the ratio of organic solvent in mobile phase, pH value, velocity of flow and the temperature of the column, the paper suggests the best chromatogram references on cordycepin including 6% acetonitrile in mobile phase, super purified solution of 0.01mol/L KH2PO4, controlling the flow velocity at 0.6ml/min, and setting the limitation of detection at 0.02.According to the optimized detecting conditions, the paper detected and compared the cordycepin contents of the cultured Cordyceps militaris fruit body and the wilding Cordyceps sinensis found in Qinghai province.2.2 optimization cordycepic acid isolation method and detectionits contents Through continuous macerating a same kind Cordyceps samples within different time, the paper made it sure that 6.5 hours is the best condition for cordycepic acid distillation. Within such a period time, the cordycepic acid will be extracted best, and the efficiency of distillation is greatest. According to the optimized distillation method, the paper distilled, detected respectively and compared the cordycepic acid contents in the cultured Cordyceps militaris fruit body and the wilding Cordyceps sinensis found in Qinghai province.2.3 Isolation cordycepic polysaccharide and detection its contents Firstly, determine the nature of cordycepic polysaccharide in the cultured Cordyceps militaris fruit body and the wilding Cordyceps sinensis found in Qinghai province, by Fehling's reaction and FTIR (Fourier transform infrared spectroscopy). Then measured the cordycepic polysaccharide contents of them respectively by Colorimetry of anthrone-sulfate.2.4 Isolation Amino acid and detection its contentsFirstly hydrolyzed the dry samples of the cultured Cordyceps militaris fruit body and the wilding Cordyceps sinensis found in Qinghai province. Then measured the cordycepic Amino acid compose and contents of them respectively by amino acid auto analyzer. Firstly used statistic method--orthogonal design in five factors and four levels to research the culture condition of Cordyceps militaris fruit body ferment, and attained the best combination of culture conditions. Thus sharply shortened the time of fruit body bourgeons and growth, and accordingly laid a solid foundation of Cordyceps militaris industrialized mass production. Chose the wilding Cordyceps sinensis found in Qinghai province as standard experimental materials, reviewed all the current detection methods of Cordyceps' active components-- cordycepin and cordycepic acid, finally attainted the optimized isolation and detection methods of them. Consequently, established a unitive measured standards, and then detected the active components contents of the two different types Cordyceps and those of their different parts. Adopted the optimized methods, at the same conditions, the paper detected the active components in...