色谱

色谱

• 技术与应用 • 上一篇    

三尖杉悬浮培养细胞中hinokiol的定量分析

徐晓辉1,2, 张卫1,3*, 姚长洪1,2, 曹旭鹏1, 薛松1*   

  1. 1. 中国科学院大连化学物理研究所海洋生物产品工程组, 辽宁 大连 116023; 2. 中国科学院研究生院, 北京 100049; 3. 澳大利亚Flinders大学医学院, 海洋生物过程与生物产品研究中心和医学生物技术部, 阿德莱德SA 5042
  • 收稿日期:2011-03-02 修回日期:2011-04-29 出版日期:2011-06-28 发布日期:2011-07-25
  • 通讯作者: 张卫,博士,教授. E-mail: Wei.Zhang@flinders.edu.au.薛 松,博士,研究员. Tel: (0411)84379069,
  • 基金资助:

    国家自然科学基金项目(No. 20676130).

Quantitative analysis of hinokiol from cell suspension cultures of Cephalotaxus fortunei

XU Xiaohui1,2, ZHANG Wei1,3*, YAO Changhong1,2, CAO Xupeng1, XUE Song1*   

  1. 1. Marine Bioproducts Engineering Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China; 2. Graduate University of Chinese Academy of Sciences, Beijing 100049, China; 3. Flinders Centre for Marine Bioprocessing and Bioproducts and Department of Medical Biotechnology, School of Medicine, Flinders University, Adelaide, SA 5042, Australia
  • Received:2011-03-02 Revised:2011-04-29 Online:2011-06-28 Published:2011-07-25

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摘要： 建立了高效液相色谱分析三尖杉悬浮培养细胞中次生代谢产物的方法,实现了次生代谢产物的分离以及hinokiol的定量分析。样品经甲醇提取后,再用氨水-氯仿萃取。采用Apollo C18色谱柱(250 mm×4.6 mm, 5 μm)进行分离,流动相为甲醇-水,梯度洗脱,柱温为30 ℃,检测波长为290 nm,流速为1 mL/min。在0.0125~0.2 g/L范围内,hinokiol的色谱峰面积与质量浓度之间具有良好的线性关系。采用该方法测定了实际样品中hinokiol的含量,并进行了3个水平的加标回收试验,其回收率为87.2%～94.7%,相对标准偏差(n=3)为0.9%~4.2%。该方法可靠、重现性好,适合对植物培养细胞中的hinokiol进行分析。

关键词: hinokiol, 高效液相色谱, 三尖杉, 松香烷二萜, 悬浮培养细胞

Abstract: A high performance liquid chromatography (HPLC) method was developed for the separation of secondary metabolites and quantitative analysis of hinokiol from cell suspension cultures of Cephalotaxus fortunei. The samples were prepared by extraction using methanol followed by partitioning between ammonium hydroxide and chloroform. The HPLC separation was achieved on an Apollo C18 column (250 mm×4.6 mm, 5 μm) with gradient elution using methanol and water at 1 mL/min and 30 ℃. The detection was carried out at 290 nm. A good linear correlation between the hinokiol peak area and mass concentration was observed over the mass concentration range of 0.0125~0.2 g/L. The proposed method was applied to the determination of hinokiol in the actual samples with recoveries of 87.2%~94.7% and with the relative standard deviations of 0.9%~4.2%. This method is reliable and reproducible and is suitable for the analysis of hinokiol in plant cell cultures.

Key words: abietane diterpenoid, cell suspension cultures, Cephalotaxus fortunei, hinokiol, high performance liquid chromatography (HPLC)