高速逆流色谱法用于纵条纹炭角菌中过氧麦角甾醇的分离纯化

黄毅; 雷传文; 宋航; 杨智芹; 袁小红; 贺新生

doi:10.13386/j.issn1002-0306.2016.17.043

高速逆流色谱法用于纵条纹炭角菌中过氧麦角甾醇的分离纯化

1. 西南科技大学生命科学与工程学院
2. 四川大学化学工程学院

基金项目:

国家自然科学基金(21272189)；四川省苗子工程项目(2015RZ0015)；西南科技大学创新基金(CX14-041)；西南科技大学研究生创新基金(15ycx090)；

详细信息

作者简介:
黄毅(1979-),男,博士,讲师,研究方向:真菌天然产物开发,E-mail:huangyi@swust.edu.cn。;

中图分类号: O652.63;O629.21
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出版历程
- 收稿日期: 2016-03-17

Isolation and purification of ergosterol peroxide from Xylaria Striata by high-speed counter-current chromatography

School of Life Science and Engineering,Southwest University of Science and Technology
Department of Pharmaceutical and Biological Engineering,Sichuan University

摘要

摘要: 过氧麦角甾醇生物活性多样,具有较大开发价值,但传统柱色谱分离手段步骤多效率低,本文建立了高速逆流色谱从纵条纹炭角菌中高效快速制备过氧麦角甾醇的方法。通过高效薄层色谱和高效液相色谱测定分配系数,再利用高速逆流色谱考查了多个溶剂体系的分离效果,确定最佳溶剂体系为:正己烷-乙酸乙酯-乙醇-水（体积比为3∶1∶2∶0.8）。以上相为固定相,下相为流动相,转速为800 r/min（正转）,流速为2 m L/min,检测波长为220 nm。制备所得过氧麦角甾醇经13C-NMR和1H-NMR鉴定,并经高效液相色谱分析纯度达到96.05%（峰面积归一化法）。经DPPH体外抗氧化活性测试,发现该化合物在0.2 mg/m L时,对DPPH自由基清除为28.36%,远低于阳性对照抗坏血酸（93.41%）,且量价关系也不明显,表明其DPPH自由基清除能力较弱。该方法制备过氧麦角甾醇简便、快速,所得产物纯度高,可用于纵条纹炭角菌中该化合物的分离。
- 过氧麦角甾醇 /
- 高速逆流色谱 /
- 纵条纹炭角菌 /
- 抗氧化活性
Abstract: Due to the various bio- activities,Ergosterol peroxide presents great application value. The column chromatography is the most commonly methods used in separation of this compound. However,sample loss caused by irreversible adsorption is very uneconomical and biggest challenges in industrial application.In present work,an effective and rapid high- speed counter- current chromatography( HSCCC) method for isolating and purifying ergosterol peroxide from Xylaria Striata has been developed.After using HPTLC and HPLC to determinate the partition coefficient,the isolation efficiency of different solvent systems was observed.The result showed that the solvent system composed of n- hexane,ethyl acetate,ethanol and water( 3 ∶ 1 ∶ 2 ∶ 0.8,v ∶ v ∶ v ∶ v) was the best,the lower phase was used as the mobile phase and performed at a flow rate of 2 m L / min,while the apparatus rotated at 800 r / min,and detected at 220 nm.The prepared ergosterol peroxide was identified by13C-NMR and1H-NMR. Its purity was 96.05% analyzed by high performance liquid chromatography.And the antioxidant potential of ergosterol peroxide was also investigated by the method of DPPH radical scavenging activity. Results revealed that ergosterol peroxide showed weaker DPPH radical scavenging activity( 28.36%) at the concentration of 0.2 mg / m L compared with standard compound ascorbic acid( 93.41%).The dose dependent was not observed also explained its weak DPPH radical scavenging activity.The established method was quite simple,fast,and suitable for the large- scale isolation and separation of ergosterol peroxide from Xylaria Striata.
- Ergosterol peroxide /
- high-speed counter-current chromatography /
- Xylaria Striata /
- antioxidant potential

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