Liquid chromatography ion trap mass spectrometric analysis of oligosaccharides using permethylated derivatives
Liquid chromatography ion trap mass spectrometric analysis of oligosaccharides using permethylated derivatives
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DOI：
10.1002/rcm.230
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发表时间：
2001-01-01
影响因子：
2
通讯作者：
Vouros, P
中科院分区：
文献类型：
--
作者：
Delaney, J;Vouros, P
将反相液相色谱与离子陷阱质谱仪的多级质量分析能力相结合，用于全甲基化低聚糖混合物的表征。新方法被用来分离未标记的过甲基化麦芽低聚物梯、2-氨基苯甲酰胺标记的麦芽低聚物梯、2AB标记的双(B)、三(T)和四天线(Q)标准的复杂混合物以及来自不同唾液酸(S)含量的可溶性CD的重组糖蛋白碳水化合物的混合物。利用反相高效液相色谱法，根据结构分离了包括α和β异构体在内的过甲基化混合物组分。全甲基化前用2-氨基苯甲酰胺荧光标记用于离线方法开发，但不一定是质谱分析所必需的，因为全甲基化本身就改善了分子的电离和碎裂特性。在MS2中确定了过甲基化衍生物的天线组成，其中Y-和B-离子系列的碎裂模式占主导地位，然后在MS3中进行了进一步的评估，这提供了从A和X交叉环碎裂获得的分支的额外信息。版权所有(C)2001 John Wiley&Sons，Ltd.
Reversed phase liquid chromatography was combined with the multiple stage mass analysis capability of an ion trap mass spectrometer for the characterization of permethylated oligosaccharide mixtures. The new method was used to separate the components of an unlabeled permethylated maltooligomer ladder, a 2-aminobenzamide-labeled (2-AB) maltooligomer ladder, a complex mixture of 2AB-labeled bi- (B), tri- (T), and tetraantennary (Q) standards, and a mixture of recombinant glycoprotein carbohydrates from soluble CD4 with varying sialic acid (S) content. Using reversed phase HPLC, permethylated mixture components including alpha and beta anomers were separated based on their structures. Fluorescent labeling with 2-aminobenzamide prior to permethylation was employed for off-line method development, but was not necessarily required for mass spectral analysis, as permethylation alone improved the ionization and fragmentation characteristics of the molecules. Antennae composition of permethylated derivatives was determined in MS2 where the fragmentation patterns of the Y- and B-ion series predominated, and then further evaluated in MS3, which provided additional information on branching obtained from A and X cross-ring fragmentation. Copyright (C) 2001 John Wiley & Sons, Ltd.