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  

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

作　　者：郭文[1] 张英[1] 黄琳娟[1] 王仲孚[1] 

机构地区：[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

出　　处：《色谱》2010年第8期776-781,共6页Chinese Journal of Chromatography

基　　金：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);陕西省重点实验室重点科研计划项目(No.09JS801)

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(250mm×4.6mm,5μm),乙腈和乙酸铵水溶液为流动相(pH4.5),梯度洗脱,在254nm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。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 a-mines by NaBH3CN. Under the optimized high performance liquid chromatography （ HPLC） and electros-pray 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.

关 键 词：液相色谱-电喷雾质谱 3-氨基-9-乙基咔唑 柱前衍生 壳寡糖 

分 类 号：O658[理学—分析化学]