H. Komatsu et al., DETERMINATION OF THE MOLECULAR-WEIGHT DISTRIBUTION OF LOW-MOLECULAR-WEIGHT HEPARINS USING HIGH-PERFORMANCE GEL-PERMEATION CHROMATOGRAPHY, Biological & pharmaceutical bulletin, 16(12), 1993, pp. 1189-1193
The molecular-weight distribution of commercially available low-molecu
lar-weight heparins (LMWHs) was estimated by high-performance gel perm
eation chromatography (HPGPC) using a LMWH molecular-weight calibrant,
supplied by the National Institute for Biological Standards and Contr
ol for the calibration of columns. This calibrant is a mixture of many
polysaccharides with a disaccharide unit prepared by a random depolym
erization of unfractionated heparin, and the ends of the fragments hav
e a UV absorption. The ratio of the refractive index to the UV absorpt
ion is a relative measure of molecular weight. Various molecular-weigh
t parameters such as weight-, number- or z-average molecular-weights,
polydispersity and relative distributions of high, middle and low mole
cular-weight components were estimated, indicating that LMWHs have a w
ide variation in molecular-weight distribution. This variation was asc
ribed to the difference in preparation methods from unfractionated hep
arins.