Development and qualification of a size exclusion chromatography coupled with multiangle light scattering method for molecular weight determination of unfractionated heparin.
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.
Beirne
John
J
Scientific Protein Laboratories LLC, 700 East Main Street, Waunakee, WI 53597, USA.
Truchan
Hilary
H
Rao
Lin
L
D000925:Anticoagulants / Q000737:chemistry*; D002850:Chromatography, Gel / Q000379:methods*; D004340:Drug Contamination; D006493:Heparin / Q000737:chemistry*; D008027:Light; D008970:Molecular Weight; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity
D000925:Anticoagulants; D006493:Heparin