Disaccharide Analysis of Glycosaminoglycans Using Hydrophilic Interaction Chromatography and Mass Spectrometry

被引：66

作者：

Gill, Vanessa Leah ^{[1
]}

Aich, Udayanath ^{[2
]}

Rao, Srinivasa ^{[2
]}

Pohl, Chris ^{[2
]}

Zaia, Joseph ^{[1
]}

机构：

[1] Boston Univ, Sch Med, Dept Biochem, Boston, MA 02118 USA

[2] Thermo Fisher Sci, Sunnyvale, CA USA

来源：

ANALYTICAL CHEMISTRY | 2013年 / 85卷 / 02期

关键词：

HEPARAN-SULFATE; LIQUID-CHROMATOGRAPHY; DOMAIN-STRUCTURE; MOLECULAR DIVERSITY; CHONDROITIN SULFATE; ELECTROPHORESIS; OCTASACCHARIDES; QUANTIFICATION; HYALURONAN; ROLES;

D O I：

10.1021/ac3030448

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Heparan sulfate (HS) and chondroitin sulfate/dermatan sulfate (CS/DS) glycosaminoglycans (GAGs) participate in many important biological processes. Quantitative disaccharide analysis of HS and CS/DS is essential for the characterization of GAGs and enables modeling of the GAG domain structure. Methods involving enzymatic digestion and chemical depolymerization have been developed to determine the type and location of sulfation/acetylation modifications as well as uronic acid epimerization. Enzymatic digestion generates disaccharides with Delta-4,5-unsaturation at the nonreducing end. Chemical depolymerization with nitrous acid retains the uronic acid epimerization. This work shows the use of hydrophilic interaction liquid chromatography mass spectrometry (HILIC-MS) for quantification of both enzyme-derived and nitrous acid depolymerization products for structural analysis of HS and CS/DS. This method enables biomedical researchers to determine complete disaccharide profiles on GAG samples using a single LC-MS platform.

引用

页码：1138 / 1145

页数：8

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