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1、网络出版时间:2013-01-0915:26网络出版地址:http://www.cnki.net/kcms/detail/12.1108.R.20130109.1526.005.html高速逆流色谱法分离制备蛹虫草发酵液中虫草素1,21,31,21,21,2*胡瑕,谢红旗,罗巍,夏志兰,刘东波1.湖南农业大学园艺园林学院,湖南长沙4101282.国家中医药管理局亚健康干预技术实验室,湖南长沙4101283.湖南省作物种质创新与资源利用重点实验室,湖南长沙410128摘要:目的以蛹虫草发酵液虫草素粗提物为原料,对高速逆流色谱(HSC
2、CC)分离制备虫草素条件进行研究。方法利用HPLC测定分配系数法结合分析型HSCCC对分离虫草素的溶剂体系进行筛选,确定虫草素分离的最佳溶剂体系为醋酸乙酯-正丁醇-0.5%氨水(2∶3∶5),并运用此溶剂体系,上相作为固定相,下相作为流动相,利用制备型HSCCC分离制备蛹虫草发酵液中虫草素。结果400mg虫草素粗产品通过制备型HSCCC一次分离制备获得质量分数为98.7%的虫草素产品43.8mg。结论该法效率高,操作简单,为虫草素的大量制备提供了重要参考,同时可为虫草素的药理研究提供物质基础。关键词:虫草素;高速逆流色谱;蛹虫草;
3、分配系数;物质基础中图分类号:R284.2;R286.02文献标志码:A文章编号:0253-2670(2013)05-0000-05DOI:10.7501/j.issn.0253-2670.2013.05.000IsolationandpurificationofcordycepinfromCordycepsmilitarisfermentationbrothbyhighspeedcounter-currentchromatography1,21,31,21,21,2HUXia,XIEHong-qi,LUOWei,XIAZhi-l
4、an,LIUDong-bo1.CollegeofHorticultureandLandscape,HunanAgriculturalUniversity,Changsha410128,China2.Sub-healthInterventionTechnologyLaboratory,StateAdministrationofTraditionalChineseMedicine,Changsha410128,China3.KeyLaboratoryforCropGermplasmInnovationandUtilizationofH
5、unanProvince,Changsha410128,ChinaAbstract:ObjectiveTodevelopanewmethodfortheisolationandpurificationofcordycepinbyhighspeedcounter-currentchromatography(HSCCC)withCordycepsmilitarisfermentationbrothasrawmaterial.MethodsThesolventsystemsforcordycepinseparationwereasses
6、sedandselectedbyHPLCpartitioncoefficientmethodandanalyticalHSCCC.Asolventsystemthatconsistedofethylacetate-n-butanol-0.5%ammoniawater(2:3:5)wasappliedtotheseparation.Theupperphasewasusedasthestationaryphase,whilethelowerphasewasusedasthemobilephaseforthecordycepinsepa
7、rationbyHSCCC.ResultsAhighefficiencyofHSCCCseparationwasachieved.Finally,cordycepin(43.8mg)wasobtainedfrom400mgcrudeextractofC.militarisfermentationbrothinone-stepseparationwithapurityof98.7%.ConclusionHSCCCisanefficientandsimplemethodforthelarge-scalepreparationofcor
8、dycepinfromC.militarisfermentationbroth,whichcouldprovidethematerialbasisforthepharmacologicalresearchofcordycepin.Keywords: