Capillary electrophoresis for total glycosaminoglycan analysis

Ebru Ucakturk, Chao Cai, Lingyun Li, Guoyun Li, Fuming Zhang, Robert J. Linhardt

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

A capillary zone electrophoresis-laser-induced fluorescence detection (CZE-LIF) method was developed for the simultaneous analysis of disaccharides derived from heparan sulfate, chondroitin sulfate/dermatan sulfate, hyaluronan, and keratan sulfate. Glycosaminoglycans (GAGs) were first depolymerized with the mixture of GAG lyases (heparinase I, II, III and chondroitinase ABC and chondroitinase AC II) and GAG endohydrolase (keratinase II) and the resulting disaccharides were derivatized by reductive amination with 2-aminoacridone. Nineteen fluorescently labeled disaccharides were separated using 50 mM phosphate buffer (pH 3.3) under reversed polarity at 25 kV. Using these conditions, all the disaccharides examined were baseline separated in less then 25 min. This CZE-LIF method gave good reproducibility for both migration time (≤1.03 % for intraday and ≤4.4 % for interday) and the peak area values (≤5.6 % for intra- and ≤8.69 % for interday). This CZE-LIF method was used for profiling and quantification of GAG derivative disaccharides in bovine cornea. The results show that the current CZE-LIF method offers fast, simple, sensitive, reproducible determination of disaccharides derived from total GAGs in a single run. [Figure not available: see fulltext.]

Original languageEnglish
Pages (from-to)4617-4626
Number of pages10
JournalAnalytical and Bioanalytical Chemistry
Volume406
Issue number19
DOIs
StatePublished - Jul 2014
Externally publishedYes

Keywords

  • Biopolymers
  • Capillary electrophoresis
  • Enzymes

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