Protocol

Monosaccharide Analysis by Methanolysis

This protocol was adapted from "Monosaccharide Analysis by Methanolysis," 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.

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INTRODUCTION

Methanolysis allows rapid determination of monosaccharide composition of glycans, glycoproteins, and proteoglycans by cleaving glycosidic bonds to produce methyl glycosides. The free-hydroxyl groups on the glycosides are derivatized with trimethylsilyl (TMS) groups to make the compounds volatile before they are analyzed by gas chromatography (GC) or gas chromatography coupled with mass spectrometry (GC-MS). The method gives more complex chromatograms than the corresponding alditol acetate method (see Monosaccharide Composition Analysis: Alditol Acetates), because up to four methyl glycosides can be produced from each monosaccharide. However, the method is easier to perform.

MATERIALS

Reagents

Acetic anhydride

t-Butyl alcohol . . . [Full Text of this Article]

Equipment

METHOD