天然产物研究与开发 ›› 2018, Vol. 30 ›› Issue (11): 1858-1862.doi: 10.16333/j.1001-6880.2018.11.002

• 研究论文 • 上一篇    下一篇

胶红酵母虾青素纯品提取及分离纯化方法优化

黄开森,廖志赢,徐春厚*,谢为天   

  1. 广东海洋大学农学院,湛江 524088
  • 出版日期:2018-12-03 发布日期:2018-12-03
  • 基金资助:

    广东省科技计划(2017A020217007);广东省科技计划(2013B020307013);湛江市科技计划(2015B01013)

Extraction,Separation and Purification Methods of Astaxanthin Pure Product from Rhodotorula mucilaginosa

HUANG Kai-sen, LIAO Zhi-ying, XU Chun-hou*, XIE Wei-tian   

  1. College of Agriculture,Guangdong Ocean University,Zhanjiang 524088,China
  • Online:2018-12-03 Published:2018-12-03

摘要: 以胶红酵母ZTHY2作为提取虾青素的出发菌株,通过对其破壁方法、浸提溶剂、皂化方法及提取条件的优化,筛选出胶红酵母虾青素纯品的提取方法。用酸热法、冻融超声波裂解法与二甲基亚砜无水乙醇法等法处理干菌体细胞,用KOH结合甲醇A、甲醇B与乙醇法分别对虾青素粗提液进行皂化处理,分别使用六种展开剂,通过薄层层析鉴定游离的虾青素,用200~300目硅胶对游离的虾青素进行柱层析,获得其纯品,最后用高效液相色谱仪测定虾青素纯品的含量。实验结果表明:胶红酵母ZTHY2干菌体细胞的最佳破壁方法是酸热法,浸提溶剂为乙酸乙酯+乙醇(2∶1)溶液,皂化液为20 g/L KOH的乙醇溶液,皂化条件是40 ℃、30 min;最佳展开剂和淋洗剂是正己烷+丙酮(5∶2)溶液;高效液相色谱仪测定的游离虾青素比率为35%,提取率为96.8%。

关键词: 胶红酵母, 虾青素, 提取方法, 酸热法

Abstract: In order to screen out a method of extracting pure astaxanthin from Rhodotorula mucilaginosa through optimizing wall breaking method,extraction solvent,saponification method and extraction conditions, Rhodotorula mucilaginosa ZTHY2 was selected as the starting strains for astaxanthin extraction.Initially,dry bacterial cells were processed through acid heat method,freeze thawing ultrasonic cleavage method and dimethyl sulfoxide anhydrous ethanol method.Three saponification methods of astaxanthin crude extract,including methanol A method mixed with KOH,methanol B method mixed with KOH and ethanol mixed method with KOH,were adopted with six developing solvents separately.Subsequently,the free astaxanthin was identified by Thin-layer chromatography implemented by column chromatography with 200-300 mesh silica gel to obtain its purity.Ultimately,astaxanthin pure product was determined by High performance liquid chromatograph (HPLC).Results showed that the best breaking wall method of Rhodotorula mucilaginosa ZTHY2 dry cell is acid heat method,and the extraction solvent is a mixture of ethyl acetate and ethanol (2∶1).Furthermore,the saponification solution is ethanol solution of 20 g/L potassium hydroxide and the saponification conditions is 40 ℃,30 min.The adopted spreading agent and eluent is a mixture of n-hexane and acetone (5∶2).The free astaxanthin ratio determined by HPLC was 35%,and the extraction rate was 96.8%.

Key words: Rhodotorula mucilaginosa, astaxanthin, extraction method, acid thermal process

中图分类号: 

R284.2