Capillary electrophoresis for the analysis of glycosaminoglycan-derived disaccharides
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Authors
Chang, Yuqing
Yang, Bo
Weyers, Amanda
Linhardt, Robert J.
Issue Date
2013-01-01
Type
Book chapter
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Alternative Title
Abstract
A quantitative and highly sensitive method for the analysis of glycosaminoglycan (GAG)-derived disaccharides is presented that relies on capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection. This method enables complete separation of seventeen GAG-derived disaccharides in a single run. Unsaturated disaccharides were derivatized with 2-aminoacridone (AMAC) to improve sensitivity. The limit of detection was at the attomole level and about 100-fold more sensitive than traditional CE-ultraviolet detection. A CE separation timetable was developed to achieve complete resolution and shorten analysis time. The RSD of migration time and peak areas at both low and high concentrations of unsaturated disaccharides are all less than 2.7% and 3.2%, respectively, demonstrating that this is a reproducible method. This analysis was successfully applied to cultured Chinese hamster ovary cell samples for determination of GAG disaccharides. The current method simplifies GAG extraction steps, and reduces inaccuracy in calculating ratios of heparin/heparan sulfate to chondroitin sulfate/dermatan sulfate, resulting from the separate analyses of a single sample.
Description
Chapter 7, In Capillary Electrophoresis of Biomolecules, Methods in Molecular Biology, 984:67-77, F. Maccari, N. Volpi eds., Humana Press
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Full Citation
Capillary electrophoresis for the analysis of glycosaminoglycan-derived disaccharides, Y. Chang, B. Yang, A. Weyers, R. J. Linhardt, Chapter 7, In Capillary Electrophoresis of Biomolecules, Methods in Molecular Biology, 984:67-77, F. Maccari, N. Volpi eds., Humana Press, 2013.
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ISSN
10643745