Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4246

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Monosaccharide Composition Analysis: Alditol Acetates

David Oxley, Graeme Currie, and Antony Bacic

This protocol was adapted from "Monosaccharide Composition Analysis: Alditol Acetates," contributed by David Oxley, Graeme Currie, and Antony Bacic, Chapter 25, in Purifying Proteins for Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2004.

The first 100 words of the full text of this article appear below.


INTRODUCTION

This protocol describes how to determine the monosaccharide composition of glycans, glycoproteins, or proteoglycans by hydrolyzing the sample to monosaccharides and converting them to alditols, then performing acetylation to make them volatile compounds and analysis by gas chromatography (GC) or gas chromatography coupled with mass spectrometry (GC-MS). It requires more work than the corresponding methanolyis procedure (see Monosaccharide Analysis by Methanolysis), but it gives good results for complex samples.


MATERIALS

Reagents

caution caution Acetic acid (5% in methanol)

caution Acetic anhydride (100%)

caution Dichloromethane (DCM) (100%)

caution Glacial acetic acid

myo-inositol (Sigma)

Glycoprotein, proteoglycan, or glycan sample of interest, vacuum-dried (1-5 µg)

caution Methanol

caution NH4OH . . . [Full Text of this Article]

Equipment


METHOD


TROUBLESHOOTING


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Related Protocol

Monosaccharide Analysis by Methanolysis
David Oxley, Graeme Currie, and Antony Bacic
CSH Protocols 2006: 4247. [Extract] [Full Text]