Determination of the Molecular-Weight Distribution of Low-Molecular-Weight Heparins Using High-Performance Gel Permeation Chromatography

• Published in: Biological & pharmaceutical bulletin Vol. 16; no. 12; pp. 1189 - 1193
• Main Authors: TAKAHATA, Tomoko, ISHIMITSU, Susumu, TANAKA, Masakazu, OKADA, Satoshi, KOMATSU, Hiroaki
• Format: Journal Article
• Language: English
• Published: Tokyo The Pharmaceutical Society of Japan 1993; Maruzen; Japan Science and Technology Agency
• Subjects: Analysis; Biological and medical sciences; Calibration; Chromatography, Gel - methods; General pharmacology; heparin; Heparin, Low-Molecular-Weight - chemistry; Heparin, Low-Molecular-Weight - isolation & purification; high-performance gel permeation chromatography; low-molecular-weight heparin; Medical sciences; molecuar-weight distribution; Molecular Weight; Pharmacology. Drug treatments; polysaccharide; Spectrophotometry, Ultraviolet; High performance; Low; Glycosaminoglycan; Qualitative analysis; Anticoagulant; Commercial form; Heparin; Molecular weight; Gel permeation chromatography
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.16.1189

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 molecuar-weight distribution. This variation was ascribed to the difference in preparation methods from unfractionated heparins.