Development and qualification of a size exclusion chromatography coupled with multiangle light scattering method for molecular weight determination of unfractionated heparin

John Beirne, Hilary Truchan, Lin Rao*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

The molecular weight of unfractionated heparin was determined by size exclusion chromatography (SEC) coupled with multiangle light scattering (MALS) detection. The SEC/MALS method determines absolute molecular weight directly from the angular dependence of scattered light intensity as a function of concentration and does not rely on molecular weight standards for column calibration. The SEC/MALS method developed at Scientific Protein Laboratories was qualified in terms of specificity, precision, robustness, and accuracy. By eliminating the requirement of well-characterized molecular weight standards derived from heparin, the present procedure represents a clear improvement over the column calibration methods used in molecular weight determination. The SEC/MALS method is suitable for routine quality control of unfractionated heparin.

Original languageEnglish (US)
Pages (from-to)717-725
Number of pages9
JournalAnalytical and Bioanalytical Chemistry
Volume399
Issue number2
DOIs
StatePublished - Jan 2011
Externally publishedYes

Keywords

  • dn/dc
  • Heparin
  • Molecular weight and molecular weight distribution
  • SEC/MALS
  • Second virial coefficient A

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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