Simultaneous determination of heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan glycosaminoglycan disaccharides by high-performance liquid chromatography using a reverse-phase column with adamantyl groups

J Chromatogr A. 2023 Jan 25:1689:463748. doi: 10.1016/j.chroma.2022.463748. Epub 2022 Dec 23.

Abstract

Glycosaminoglycans (GAGs), which are one of the major components of proteoglycans, play a pivotal role in physiological processes such as signal transduction, cell adhesion, growth, and differentiation. Characterization of GAGs is challenging due to the tremendous structural diversity of heteropolysaccharides with numerous sulfate or carboxyl groups. In this present study, we examined the analysis of 2-aminobenzamide (2-AB) labeled GAG disaccharides by high-performance liquid chromatography (HPLC) using a reverse-phase (RP)-column with adamantyl groups. Under the analytical conditions, 17 types of 2-AB labeled GAG disaccharides derived from heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan were sequentially separated in a single analysis. The analysis time was fast with high retention time reproducibility. Moreover, the RP-HPLC column with adamantyl groups allowed the quantification of GAGs in various biological samples, such as serum, cultured cells, and culture medium.

Keywords: Adamantly group; RP-HPLC; chondroitin/dermatan sulfate detection; heparan sulfate detection; Δ-Disaccharide GAG analysis.

MeSH terms

  • Chondroitin Sulfates* / chemistry
  • Chromatography, High Pressure Liquid / methods
  • Dermatan Sulfate / analysis
  • Dermatan Sulfate / chemistry
  • Dermatan Sulfate / metabolism
  • Disaccharides / chemistry
  • Glycosaminoglycans* / chemistry
  • Heparitin Sulfate / analysis
  • Hyaluronic Acid / analysis
  • Hyaluronic Acid / chemistry
  • Reproducibility of Results

Substances

  • Glycosaminoglycans
  • Chondroitin Sulfates
  • Hyaluronic Acid
  • Dermatan Sulfate
  • Disaccharides
  • Heparitin Sulfate