色谱

色谱 ›› 2012, Vol. 30 ›› Issue (03): 304-308.DOI: 10.3724/SP.J.1123.2011.11015

• 研究论文 • 上一篇    下一篇

毛细管电泳法分析藏红花植物细胞多糖中单糖组成

马海宁, 华玉娟, 屠春燕*, 袁丽红, 韦萍   

  1. 南京工业大学生物与制药工程学院, 江苏 南京 210009
  • 收稿日期:2011-11-09 修回日期:2011-12-30 出版日期:2012-03-28 发布日期:2012-05-02
  • 通讯作者: 屠春燕,硕士,研究员,主要研究方向为仪器分析.
  • 基金资助:

    国家重点基础研究发展计划项目(2009CB724700)

Analysis of monosaccharides in the saffron corm glycoconjugate by capillary electrophoresis

MA Haining, HUA Yujuan, TU Chunyan*, YUAN Lihong, WEI Ping   

  1. College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China
  • Received:2011-11-09 Revised:2011-12-30 Online:2012-03-28 Published:2012-05-02

PDF

367

被引次数

32

摘要：

以对甲氧基苯胺为衍生试剂,采用毛细管电泳法分析了藏红花植物细胞多糖中的单糖组成。对衍生条件进行了优化,并对毛细管分离条件进行了系统的研究。衍生反应在醋酸含量9.5%(v/v)、80 ℃下反应2 h的衍生产率最大,衍生产物紫外检测波长234 nm。在优化的毛细管电泳分离条件(未涂层熔融石英毛细管柱(60 cm(有效长度50 cm)×50 μm),柱温25 ℃,电压20 kV,使用350 mmol/L硼酸电解液(pH 10.21),压力(3.4475 kPa)进样5 s)下,基线分离了11种结构相近的醛糖(来苏糖、木糖、核糖、葡萄糖、甘露糖、半乳糖、鼠李糖、纤维二糖、麦芽糖、乳糖)、酮糖(果糖)的衍生产物。应用该方法定量检测了藏红花植物细胞多糖水解物中糖的成分,各糖的回收率为94.3%～105.4%,相对标准偏差为3.3%～4.6%。

关键词: 藏红花, 单糖, 对甲氧基苯胺, 多糖, 毛细管电泳, 酮糖, 柱前衍生

Abstract:

The monosaccharides in the saffron corm glycoconjugate were separated by capillary electrophoresis (CE) coupled with pre-column derivatization. 4-Methoxyaniline was used as derivatization reagent. The derivatization and CE separation conditions were investigated. The ultraviolet detection wavelength was 234 nm. The maximum yield of this derivatization reaction was obtained under the presence of 9.5% (v/v) acetic acid at 80 ℃ for 2 h. An uncoated fused-silica capillary of 50 μm i.d. and 50/60 cm length (effective length/total length) was employed, and a pressure injection (3.4475 kPa, 5 s) was applied. The baseline separation of 11 monosaccharides and disaccharides (lyxose, xylose, ribose, glucose, mannose, galactose, rhamnose, cellobiose, maltose, lactose, fructose) was reached at 25 ℃, 20 kV of separation voltage and with 350 mmol/L boric acid (pH 10.21) as running buffer. The developed method has been successfully applied to quantitatively determine the components of saffron corm glycoconjugate, and the results showed that the recovery of each monosaccharide was in the range of 94.3%~105.4%, the relative standard deviation was 3.3%~4.6%.

Key words: 4-methoxyaniline, glycoconjugate, ketoses, monosaccharides, pre-column derivatization, saffron, capillary electrophoresis (CE)