正交设计优选蛹草拟青霉中虫草素和虫草多糖的提取工艺

摘要

目的：采用正交设计优化蛹草拟青霉菌丝体中虫草素和虫草多糖的提取工艺。方法：建立高效液相色谱测定指标成分虫草素含量的方法，采用正交试验法考察醇浓度、加醇量、提取次数与时间的组合3个因素对虫草素乙醇提取率的影响。醇提后残渣以水提取多糖，采用苯酚-浓硫酸比色法在490 nm 处测定虫草多糖的含量，以单因素法筛选多糖提取工艺。结果：HPLC 色谱条件下精密度的 RSD 为0.30％，稳定性的 RSD 为1.27％，重复性的RSD 为2.93％。以乙醇浓度为90％，加醇量为10倍量，提取3次，每次1 h 虫草素的提取率最高；而以乙醇浓度为95％，加醇量为6倍量，提取3次，每次1 h 提取虫草素后的残渣再以水提取，虫草多糖得率最高。结论：所优化的提取工艺虫草素和虫草多糖提取率均较高，采用的含量测定方法稳定可靠。%Objective:To optimize the extraction processes of cordycepin and cordycepic polysaccharides from mycelia of Paecilomyces militaris by using Orthogonal design.Methods:A HPLC method was established to determine the content of cordycepin which was taken as indexes,and orthogonal test method was used to investigate three factors of alcohol concentration,alcohol volume ratio,and the combination of extraction times with time to influence on the extraction efficiency of cordycepin.Detection was performed at wavelength of 260 nm.The column temperature was 30 ℃ and the injection volume was 1 0 μL.After extracted by alcohol,the residue of mycelia was dealt with water to extracted cordycepic polysaccharides, and colorimetric method by using phenol—concentrated sulfuric acid as chromogenic agent was used to determine the content of cordycepic polysaccharides in the extract under 490 nm.Results:Under the condition of HPLC,the precision of RSD was 0.30%,the stability of RSD was 1 .27% and the RSD of reproducibility was 2.93%.The results showed the extract ratio of cordycepin was the highest with the process parameters of 90% alcohol,1 0 times of alcohol volume ratio,3 times with 1 h for every time.And the water extract ratio of Cordyceps polysaccharide in the mycelia residue was the highest after cordycepin was extracted from mycelia with 95% alcohol,6 times of alcohol volume ratio,3 times,with 1 h each time.Conclusion:The extraction efficiency of cordycepin and Cordyceps Polysaccharide was high under the optimum process,and the methods of the content determination were stabile and reliable.