Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Editor

Personal info

View

About

Terms

Javascript disabled

Please enable Javascript support in your browser to use this application.

Instructions on how to enable JavaScript on different browsers can be found here: http://www.google.com/support/bin/answer.py?answer=23852.

[edit this page]

Fu, D. et al.

Monosaccharide composition analysis of oligosaccharides and glycoproteins by high-performance liquid chromatography.

A simple and sensitive high-performance liquid chromatography (HPLC)-based method for complete monosaccharide composition analysis of oligosaccharides and glycoproteins is described. In this method, an oligosaccharide or glycoprotein is first hydrolyzed using an optimized method to give the constituent monosaccharides, which are subsequently labeled with 1-phenyl-3-methyl-5-pyrazolone (PMP) as previously described by Honda et al. (Anal, Biochem. 180, 351-357, (1989)). The labeled monosaccharides are separated by reverse-phase HPLC using a column developed especially for this purpose, monitored by uv absorbance at 245 nm, and quantitated by their integration values relative to standards. Sialic acids are acid-labile keto-sugars. They are, therefore, released with neuraminidases or by mild acid hydrolysis and then converted with neuraminic acid aldolase to their corresponding mannosamine derivatives, which are then PMP-labeled, separated, and quantitated as described above. Individual sialic acids including N-acetyl and N-glycolyl neuraminic acids are well resolved and quantitated by this method. This method has proven to be highly sensitive, requiring only 1 pmol for reliable detection. Quantitative analysis of neutral and amino sugars from both oligosaccharide and glycoprotein samples can be achieved using one acid hydrolysis and a set of equal molar monosaccharide standards. Similarly, quantitation of sialic acids works equally well with both free oligosaccharide and glycoprotein samples. Monosaccharide compositions of oligosaccharides and glycoproteins determined by this method were found to be highly accurate.[1]

References

Monosaccharide composition analysis of oligosaccharides and glycoproteins by high-performance liquid chromatography. Fu, D., O'Neill, R.A. Anal. Biochem. (1995) [Pubmed]

Annotations and hyperlinks in this abstract are from individual authors of WikiGenes or automatically generated by the WikiGenes Data Mining Engine. The abstract is from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.About WikiGenes Open Access Licence Privacy Policy Terms of Use apsburg

The world's first wiki where authorship really matters (Nature Genetics, 2008). Due credit and reputation for authors. Imagine a global collaborative knowledge base for original thoughts. Search thousands of articles and collaborate with scientists around the globe.