Table of Contents

A practical guide to structural analysis of carbohydrates

Introduction

This guide is a collection of standardized procedures used at Stockholm University in the laboratory of Prof. Per-Erik Jansson (now at Karolinska Institutet). The original manuscript, excluding figures, was translated to HTML by Roland Stenutz and to dokuwiki by Magnus Lundborg. The procedures are maintained in the research group of Prof. Göran Widmalm.
There are still many omissions and errors so be careful when using it and do not rely its accuracy.
Some of the reagents reffered to are highly toxic and/or corrosive and must be handled with extreme care. In most cases normal laboratory practice will suffice (fume hood, lab coat, safety goggles).
Corrections and additions are most welcome and should be addressed to Jonas Ståhle, a member of the research group of Prof. Göran Widmalm.


If you find this guide useful please cite it as:
R. Stenutz, P.-E. Jansson and G. Widmalm; “A practical guide to structural analysis of carbohydrates”
http://www.organ.su.se/gw/doku.php?id=sop:index and date of access.


Component analysis

  1. Sugar analysis
  2. Methylation analysis using partially methylated alditol acetates (PMAAs)
    1. - with methyl triflate
  3. Determination of absolute configuration using butyl glycosides

Specific degradations

  1. Acetolysis
  2. N-deacylation by trifluoroacetolysis, followed by deamination or hydrolysis
  3. N-deacylation by alkali in DMSO, followed by deamination or hydrolysis

Modification of polysaccharides

  1. Peracetylation
  2. N-acetylation
  3. N-deacylation
  4. O-deacylation
  5. Dephosphorylation
  6. Hydrazinolysis

Purification of polysaccharides

  1. Phenol-Chloroform-Petroleum ether extraction (PCP-extraction)
  2. Precipitation of acidic polysaccharides with cetyltrimethyl ammonium chloride (Cetavlon)
  3. Precipitation with ethanol

Preparation of reagents

Chromatography

  1. Biogel P4
  2. Sephadex G50
  3. Superdex
  4. DEAE-Sepharose
  5. CM-Sepharose

Web resources

Appendix - Carbohydrate structures