Absolute molecular weight distribution of low-molecular-weight heparins by size-exclusion chromatography with multiangle laser light scattering detection

Anal Biochem. 1997 Feb 15;245(2):231-41. doi: 10.1006/abio.1996.9984.

Abstract

The absolute molecular weight (M(r)) distribution of seven low-molecular-weight (LMW) heparin products was determined by size-exclusion chromatography (SEC) coupled with multiangle laser light scattering (MALLS) detection. The SEC/MALLS technique does not rely on relative M(r) standards for column calibration and yields absolute M(r) estimates directly from the angular dependence of scattered light intensity as a function of concentration, as formulated by light scattering theory. The SEC/MALLS method we describe is rapid, precise, and accurate. In 1 h it yields results from triplicate injections that agree well with the manufacturers' own independent analyses and that exhibit coefficients of variation of approximately 1%. By eliminating the requirement for finite quantities of highly purified, well-characterized M(r) standards derived from heparin, the present procedure represents a clear improvement over relative methods of M(r) determination. Thus, it is concluded that the SEC/MALLS method is ideally suited to routine quality control of commercial LMW-heparin products.

Publication types

  • Comparative Study

MeSH terms

  • Anticoagulants / chemistry
  • Chromatography, High Pressure Liquid / methods
  • Chromatography, Liquid / instrumentation
  • Chromatography, Liquid / methods*
  • Dalteparin / chemistry
  • Dextrans / chemistry
  • Heparin / chemistry*
  • Lasers*
  • Models, Chemical
  • Molecular Weight
  • Quality Control
  • Scattering, Radiation*

Substances

  • Anticoagulants
  • Dextrans
  • Heparin
  • Dalteparin