色谱

色谱 ›› 2010, Vol. 28 ›› Issue (08): 776-781.DOI: 10.3724/SP.J.1123.2010.00776

• 研究论文 • 上一篇    下一篇

高效液相色谱-质谱联用法分析3-氨基-9-乙基咔唑柱前衍生化壳寡糖

郭文, 张英, 黄琳娟, 王仲孚*   

  1. 西北大学生命科学学院, 西部资源生物与现代生物技术教育部重点实验室, 陕西 西安 710069
  • 收稿日期:2010-03-19 修回日期:2010-06-11 出版日期:2010-08-28 发布日期:2010-08-28
  • 通讯作者: 王仲孚,教授.
  • 基金资助:

    国家“863”计划项目(Nos. 2007AA10Z338, 2007AA091601)、教育部新世纪优秀人才支持计划(No. NCET-08-0893)和陕西省重点实验室重点科研计划项目(No. 09JS801).

High performance liquid chromatography/electrospray ionization ion-trap mass spectrometry analysis of chitooligosaccharides by pre-column derivatization with 3-amino-9-ethylcarbazole

GUO Wen, ZHANG Ying, HUANG Linjuan, WANG Zhongfu*   

  1. Key Laboratory of Ministry of Education for Western China Resource Biology and Biotechnology, Life Sciences College, Northwest University, Xi’an 710069, China
  • Received:2010-03-19 Revised:2010-06-11 Online:2010-08-28 Published:2010-08-28

PDF

673

被引次数

23

摘要： 以3-氨基-9-乙基咔唑(3-amino-9-ethylcarbazole, AEC)为衍生化试剂对壳寡糖进行柱前衍生,壳寡糖(COS)的还原端与AEC的伯氨基反应生成烯胺,再被硼氢氰化钠(NaBH3CN)还原为二级胺。采用反相C18色谱柱(250 mm×4.6 mm, 5 μm),乙腈和乙酸铵水溶液为流动相(pH 4.5),梯度洗脱,在254 nm波长处检测,建立了一套壳寡糖衍生物的高效液相色谱(HPLC)分离分析、电喷雾质谱(ESI-MS)及液相色谱-电喷雾质谱联用(LC-ESI-MS)的分析方法。该方法操作简单、灵敏度高、重复性好,在COS的组分分析、质量控制及构效关系研究方面有潜在的应用价值。

关键词: 3-氨基-9-乙基咔唑, 壳寡糖, 液相色谱-电喷雾质谱, 柱前衍生

Abstract: A pre-column derivatization method was established for chitooligosaccharides (COS) analysis, employing 3-amino-9-ethylcarbazole (AEC) as a derivatization reagent. The enamine was generated by the reaction of the reducing end of COS and the primary amine of AEC, and then reduced to secondary amines by NaBH3CN. Under the optimized high performance liquid chromatography (HPLC) and electrospray ionization ion-trap mass spectrometry (ESI-MS) conditions, the HPLC separation was carried out on an RP C18 column (250 mm×4.6 mm, 5 μm) with a gradient elution using acetonitrile and ammonium acetate as mobile phase. The derivative was successfully analyzed by HPLC, ESI-MS and on line HPLC-ESI-MS in positive-ion mode. The established method is simple, high sensitive and reproducible, which has great potential application to component analysis, quality control and structure activity relationship analysis of COS.

Key words: 3-amino-9-ethylcarbazole (AEC), chitooligosaccharides, pre-column derivatization, high performance liquid chromatography/electrospray ionization ion-trap mass spectrometry (LC-ESI-MS)

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