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High performance capillary electrophoresis method to characterize heparin and heparan sulfate disaccharides
Author(s) -
Karamanos Nikos K.,
Vanky Peter,
Tzanakakis George N.,
Hjerpe Anders
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150170217
Subject(s) - capillary electrophoresis , heparan sulfate , heparin , chromatography , chemistry , electrophoresis , sulfate , perlecan , biochemistry , organic chemistry
Abstract A rapid, sensitive and accurate high‐performance capillary electrophoresis method is described for the determination of the sulfation pattern of heparin and heparan sulfate disaccharides. The analysis, performed after enzymic degradation of the polysaccharides with heparinase and heparinases II and III in combination, yields highly UV‐absorbing Δ‐disaccharides. The separation is performed with reversed polarity using 15 m M phosphate buffer, pH 3.50. This method is superior to others since all known 12 disaccharides carrying N ‐acetylated, N ‐sulfated or unsubstituent glucosamine can be separated in a single run of 15 min. At the highest sensitivity the analysis consumes only a few femtograms of glycosaminoglycan and allows a determination of Δ‐disaccharides at the attomole level.