κ-卡拉胶寡糖的反相离子对-超高效液相色谱-四极杆-飞行时间质谱研究  

Analysis of Oligo-κ-carrageenan by Reversed Phase Ion-pair Ultra-Performance Liquid Chromatography Coupled with Electrospray Ionization-Time of Flight-Mass Spectrometry

作　　者：高洋[1] 陈海敏[1] 徐继林[1] 陈德莹[1] 严小军[1] 

机构地区：[1]宁波大学应用海洋生物技术教育部重点实验室,宁波315211

出　　处：《分析化学》2009年第11期1590-1595,共6页Chinese Journal of Analytical Chemistry

基　　金：教育部长江学者与创新团队项目(No.IRT0734);国家自然科学基金(No.30800860)资助项目

摘　　要：建立反相离子对-超高效液相色谱(RPIP-UPLC)和电喷雾离子源-四极杆-飞行时间质谱(ESI-Q-TOF-MS)联用技术快速分离鉴定硫酸寡糖的方法。以20mmol/L庚胺(pH4)为离子对试剂,25%庚胺甲酸盐纯水溶液(A)和25%庚胺甲酸盐甲醇溶液(B)为梯度洗脱溶剂,κ-卡拉胶寡糖通过BEHC18反相柱分离后,分别在正、负离子模式下进行四极杆-飞行时间质谱分析。结果表明,聚合度为3～45的κ-卡拉胶寡糖在BEH C18柱上得到很好的分离,从每一个色谱峰对应的质谱图中可以准确获得直至27糖的各寡糖结构信息,均为奇数糖,与聚丙烯凝胶电泳结果吻合。所得的寡糖断裂规律对卡拉胶寡糖的快速鉴定和结构解析具有重要意义。A method was developed to elucidate the structures of sulfated oligosaccharides through establishment of an effective reversed phase ion pair ultra-performance liquid chromatography coupled with electrospray ionization time of flight mass spectrometry（RPIP-UPLC-ESI-TOF-MS）. Heptylamine （20 mmol/L, pH 4） has been selected as the ion-pairing agent. κ-Carrageenan oligosaccharides have been separated on BEH Cls column using MeOH/H20 with 25% heptylammonium formate as eluent in linear gradient mode. Mass spectra were obtained by ESI-Q-TOF-MS in both positive and negative modes, κ-Carrageenan oligosaccharides were well separated up to pentatetrasaccharide, and ESIMS analysis for κ-carrageenan oligosaccharides up to heptacosasccharide. The results showed that all acid hydrolyzed κ-carrageenan oligosaccharides were odd sugars, which was further confirmed by polyacrylamide gel electrophoresis （PAGE）. The characteristic fragmentation pattem of ion-pair oligosaccharides in mass spectra can be applied for rapid structure identification.

关 键 词：Κ-卡拉胶 反相离子对色谱-超高效液相色谱 电喷雾离子源-四极杆-飞行时间质谱 聚丙烯凝胶电泳 

分 类 号：O657.63[理学—分析化学]