Monitoring the stability of heparin: NMR evidence for the rearrangement of sulfated iduronate in phosphate buffer

Carbohydr Polym. 2023 May 15:308:120649. doi: 10.1016/j.carbpol.2023.120649. Epub 2023 Feb 3.

Abstract

Heparin, a major anticoagulant drug, comprises a complex mixture of motifs. Heparin is isolated from natural sources while being subjected to a variety of conditions but the detailed effects of these on heparin structure have not been studied in depth. Therefore, the result of exposing heparin to a range of buffered environments, ranging pH values from 7 to 12, and temperatures of 40, 60 and 80 °C were examined. There was no evidence of significant N-desulfation or 6-O-desulfation in glucosamine residues, nor of chain scission, however, stereochemical re-arrangement of α-L-iduronate 2-O-sulfate to α-L-galacturonate residues occurred in 0.1 M phosphate buffer at pH 12/80 °C. The results confirm the relative stability of heparin in environments like those during extraction and purification processes; on the other hand, the sensitivity of heparin to pH 12 in buffered solution at high temperature is highlighted, providing an important insight for heparin manufacturers.

Keywords: Alkaline solutions; Heparin; Iduronate rearrangement; NMR; Phosphate buffer.

MeSH terms

  • Heparin* / chemistry
  • Iduronic Acid
  • Phosphates
  • Sulfates*

Substances

  • Heparin
  • Sulfates
  • iduronate 2-sulfate
  • Iduronic Acid
  • Phosphates