The efficient structure elucidation of minor components in heparin digests using microcoil NMR

John F K Limtiaco, S. Béni, Christopher J. Jones, Derek J. Langeslay, Cynthia K. Larive

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The structural complexity and microheterogeneity of the glycosaminoglycans heparin and heparan sulfate make their characterization a daunting task. The methodology described herein utilizes a combination of enzymatic digestion, size-exclusion chromatography, strong anion-exchange HPLC, reverse-phase ion-pair ultrahigh performance liquid chromatography-mass spectrometry, and microcoil NMR for the efficient sequencing of heparin-derived tetrasaccharides. The high mass sensitivity of microcoil NMR makes this technique well suited for the characterization of mass-limited samples removing a bottleneck in the analysis workflow and permitting structural characterization of minor components isolated from a heparin enzymatic digestion. Complete characterization of one tetrasulfonated, five pentasulfonated isomers and two hexasulfonated tetrasaccharide sequences is described. To our knowledge, two of the identified minor tetrasaccharides are unique, and have not been previously reported: IdoA(2S)-GlcNS(6S)-IdoA(2S)-GlcNS(6S) and ΔUA(2S)-GlcNS(6S)-IdoA-GlcNS(6S) .

Original languageEnglish
Pages (from-to)2244-2254
Number of pages11
JournalCarbohydrate Research
Volume346
Issue number14
DOIs
Publication statusPublished - Oct 18 2011

Fingerprint

Heparin
Nuclear magnetic resonance
Digestion
Heparitin Sulfate
Workflow
Glycosaminoglycans
Liquid Chromatography
Gel Chromatography
Anions
Size exclusion chromatography
Mass Spectrometry
Liquid chromatography
High Pressure Liquid Chromatography
Isomers
Ions
Mass spectrometry
O-(alpha-2-sulfamino-2-deoxy-D-glucopyranosyl)-(1-3)-L-idonic acid

Keywords

  • Heparin diastereomers
  • Hexuronic acid epimers
  • Microcoil NMR
  • Nuclear magnetic resonance
  • Unsaturated tetrasaccharide
  • UPLC-MS

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Organic Chemistry

Cite this

The efficient structure elucidation of minor components in heparin digests using microcoil NMR. / Limtiaco, John F K; Béni, S.; Jones, Christopher J.; Langeslay, Derek J.; Larive, Cynthia K.

In: Carbohydrate Research, Vol. 346, No. 14, 18.10.2011, p. 2244-2254.

Research output: Contribution to journalArticle

Limtiaco, John F K ; Béni, S. ; Jones, Christopher J. ; Langeslay, Derek J. ; Larive, Cynthia K. / The efficient structure elucidation of minor components in heparin digests using microcoil NMR. In: Carbohydrate Research. 2011 ; Vol. 346, No. 14. pp. 2244-2254.
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