Determination of the molecular-weight distribution of low-molecular-weight heparins using high-performance gel permeation chromatography.
The molecular-weight distribution of commercially available low-molecular-weight heparins (LMWHs) was estimated by high-performance gel permeation chromatography (HPGPC) using a LMWH molecular-weight calibrant, supplied by the National Institute for Biological Standards and Control for the calibration of columns. This calibrant is a mixture of many polysaccharides with a disaccharide unit prepared by a random depolymerization of unfractionated heparin, and the ends of the fragments have a UV absorption. The ratio of the refractive index to the UV absorption is a relative measure of molecular weight. Various molecular-weight parameters such as weight-, number- or z-average molecular-weights, polydispersity and relative distributions of high, middle and low molecular-weight components were estimated, indicating that LMWHs have a wide variation in molecular-weight distribution. This variation was ascribed to the difference in preparation methods from unfractionated heparins.
Komatsu
H
H
Division of Drugs, National Institute of Health Science, Osaka, Japan.
Takahata
T
T
Tanaka
M
M
Ishimitsu
S
S
Okada
S
S
D002138:Calibration; D002850:Chromatography, Gel / Q000379:methods*; D006495:Heparin, Low-Molecular-Weight / Q000737:chemistry* / Q000302:isolation & purification; D008970:Molecular Weight; D013056:Spectrophotometry, Ultraviolet
D006495:Heparin, Low-Molecular-Weight