title: Monosaccharide composition analysis of pamiteplase by anion exchange chromatography with pulsed amperometric detection.
abstract: The monosaccharides (neutral and amino sugars) of palmiteplase (recombinant modified human tissue plasminogen activator) were analyzed by high performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Since the palmiteplase formulation contains sucrose, it was removed by reverse-phase high-performance liquid chromatography (RP-HPLC) prior to analysis. Acid hydrolysis with TFA was performed at 100 degrees C for 4 h. Fucose, glucosamine, galactose and mannose were detected by HPAEC-PAD analysis after hydrolysis. The linearity range of HPAEC-PAD analysis was 20-200 pmol/ml(-1) (r > 0.999) and the RSD value for repeatability was less than 7%. The recovery of each monosaccharide spiked into samples was more than 90%. The monosaccharide composition of palmiteplase suggests that it has complex-type oligosaccharides lacking in high-mannose-type oligosaccharides.
authors.affiliation: Yamanouchi Pharmaceutical Co, Ltd, Tsukuba-shi, Japan. yokota_h@yamanouchi.co.jp
authors.forename: H
authors.initials: H
authors.lastname: Yokota
authors.forename: K
authors.initials: K
authors.lastname: Mori
authors.forename: H
authors.initials: H
authors.lastname: Yamaguchi
authors.forename: H
authors.initials: H
authors.lastname: Kaniwa
authors.forename: N
authors.initials: N
authors.lastname: Saisho
mesh_terms: D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000379:methods; D005643:Fucose / Q000032:analysis; D005690:Galactose / Q000032:analysis; D005944:Glucosamine / Q000032:analysis; D006801:Humans; D008358:Mannose / Q000032:analysis; D009005:Monosaccharides / Q000032:analysis*; D011994:Recombinant Proteins / Q000737:chemistry; D015203:Reproducibility of Results; D010959:Tissue Plasminogen Activator / Q000737:chemistry*; D014269:Trifluoroacetic Acid
chemical_list: D009005:Monosaccharides; D011994:Recombinant Proteins; D005643:Fucose; D014269:Trifluoroacetic Acid; D010959:Tissue Plasminogen Activator; C085027:pamiteplase; D005944:Glucosamine; D008358:Mannose; D005690:Galactose