journal6 ›› 2009, Vol. 30 ›› Issue (1): 79-83.

• 化学化工 • 上一篇    下一篇

气相色谱检测芦荟中海藻糖方法的建立

  

  1. (1.南京理工大学化工学院,江苏 南京 210094；2.北京工商大学化学与环境工程学院，北京 00037)
  • 出版日期:2009-01-25 发布日期:2012-04-27
  • 通讯作者: 何聪芬（1966-），女，河北石家庄人，教授，博士，主要从事应用生物技术研究.E-mail:hecf@th.btbu.edu.cn.
  • 作者简介:赵华（1965-），男，山西稷山人，北京工商大学化学与环境工程学院副教授，主要从事生物化工分离研究
  • 基金资助:

    国家863计划资助项目（2007AA100404）；北京市教委和北京基金委科技重点项目（KZ200410011006）

Establishment of GC to Detect Trehalose in Aloe

  1. (1.School of Chemical Engineering,Nanjing University of Science and Technology,Nanjing 210094,China;2.College of Chemistry and Environment Engineering,Beijing Technology and Business University,Beijing 100037,China)
  • Online:2009-01-25 Published:2012-04-27

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摘要：用1-甲基咪唑、氯化羟胺、乙酸酐将海藻糖衍生化，以气相色谱法作为定量分析手段，建立了植物组织中微量海藻糖定量检测方法.用该方法对同一种糖类衍生物进行多次分析，其特征峰保留时间误差在3 s内；能将D-葡萄糖、乳糖、蔗糖和海藻糖进行分离.海藻糖在（3.697～28.661）×10-9 g检测量范围内其相关系数为0.998 6.利用本实验建立的植物组织中微量海藻糖定量检测的方法，分别对3年生、5年生库拉索芦荟和半年生海藻糖合成酶基因转化芦荟凝胶中海藻糖的含量进行测定，结果表明：5年生库拉索芦荟凝胶中海藻糖含量是3年生的1.59倍，每10 g凝胶匀浆中分别为1.103×10-5 g和6.905×10-6 g；每10 g半年生海藻糖合成酶基因转化芦荟凝胶匀浆中海藻糖含量为1.614×10-5 g，是3年生的2.33倍.证明目的基因成功转入芦荟并已经表达.

关键词： 气相色谱, 海藻糖, 定量分析, 库拉索芦荟

Abstract: To determine the content of trehalose in transgenic Aloe by gas chromatography (GC),1-Methyl-imidazol,Hydroxylammonium chloride and Acetic anhydride were used to derivative the trehalose.The difference of the retention time among the same carbohydrate derivatives is 3s by this method.And it can separate the glucose,milk sugar,sucrose and trehalose clearly.The results showed a good linear relationship within a range from 3.697×10-9　g to 28.661×10-9　g,and the liner correlation coefficient of the trehalose is 0.998　6.The content of trehalose in the three-year,five-year aloe (Aloe vera.L) and aloe transformed by ostA were determined by GC.The results show that the trehalose of the fiveyear aloe (Aloe vera.L) is 1.59 times as great as the three-year.The content of trehalose in five-year aloe is 1.103×10-5　g per 10g,and 6.905×10-6　g per 10g in three-year aloe.The content of trehalose is 1.614×10-5　g per 10g in aloe transformed by ostA which is 2.33 times as great as the three-year aloe (Aloe vera.L).The results prove that the ostA gene from E.coli is successfully transformed to aloe and expressed.

Key words: gas chromatography, trehalose, quantificational analysis, Aloe vera.L

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