KMID : 0616120090400020109
|
|
Korean Journal of Pharmacognosy 2009 Volume.40 No. 2 p.109 ~ p.117
|
|
Analysis of Contaminated Heparins by the Combination of Glycosaminoglycan Degrading Enzymes and Nuclear Magnetic Resonance Spectroscopy
|
|
Im A-Rang
Kim Yeong-Shik Hong Chong-Hui Lee Su-Jung Lee Sun-Hee Park You-Mie
|
|
Abstract
|
|
|
We examined the purity of six heparin samples by using heparinase, chondroitinase, , and polyacrylamide gel electrophoresis. To obtain high molecular weight contaminants from heparin samples, heparinase I - digested samples were subjected to the exhaustive microcon filtration. The filtration process removed heparin-derived di- and oligosaccharides effectively. By combining chondroitinase ABC treatment and strong anion exchange - high performance liquid chromatography, the result showed all six samples contained chondroitin sulfate as a contaminant ranging from 1.3 to 14.9%. Among them, sample S3 showed the highest content of 14.9%, which was further analyzed by chondroitinase AC treatment to confirm chondroitin sulfate B (dermatan sulfate). chemical shifts of N-acetyl groups clearly suggested the existence of chondroitin sulfate B (sample S3) and oversulfated chondroitin sulfate (samples S2 and S4) as contaminants. In addition, polyacrylamide gel electrophoresis was useful for qualitative detection on the sample¡¯s purity. These results suggest that the tools of heparinase I and chondroitinase ABC in combination with NMR spectroscopy would give very useful information for investigation of heparin contaminants such as oversulfated chondroitin sulfate and dermatan sulfate in heparin samples.
|
|
KEYWORD
|
|
Heparin, Oversulfated chondroitin sulfate, Heparinase, Chondroitinase, Analysis, Dermatan sulfate
|
|
FullTexts / Linksout information
|
|
|
|
Listed journal information
|
|
|