Chromatographic methods for product-profile analysis and isolation of oligosaccharides produced by heparinase-catalyzed depolymerization of heparin

Analysis of heparin sodium by SAX/HPLC for contaminants and impurities

“…The SAX silica column gave the best separation of heparin from OSCS [18] but the retention times continually became shorter with column use and eventually after approximately 200 h of use the column would fail due to high pressure. Drifting elution times with silica based columns when using the high salt content eluent has been reported previously [19] and appears to be due to column degradation. The polymeric SAX Dionex AS11-HC column separated OSCS and dermatan sulfate from heparin under the elution conditions selected as shown in Fig.…”

Pharmacology of Heparin and Related Drugs

“…Heparin is found naturally in mast cell granules, where it interacts with histamine; this interaction has been analyzed in some structural detail and is highly ordered and specific, with the imidazolium ring of protonated histamine located between two IdoA2S residues by hydrogen bonds between the histamine NH and IdoA carboxylates (Chuang et al, 2000). Histamine alters the profile of oligosaccharides resulting from depolymerization with heparinase I (Chuang et al, 2001(Chuang et al, , 2002.…”

Characterization of a Heparan Sulfate 3-O-Sulfotransferase-5, an Enzyme Synthesizing a Tetrasulfated Disaccharide

“…Fractions were collected and the radioactivity was determined by liquid scintillation counting. To determine the molecular size of oligosaccharides, gel filtration HPLC was performed on a Superdex TM Peptide HR10/30 column (1 ϫ 30 cm, ϫ 2 columns in series) as described (39). The column was equilibrated with 0.2 M NaCl at a flow rate of 0.8 ml/min, at room temperature.…”