Monosaccharide composition analysis of pamiteplase by anion exchange chromatography with pulsed amperometric detection.
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.
Yokota
H
H
Yamanouchi Pharmaceutical Co, Ltd, Tsukuba-shi, Japan. yokota_h@yamanouchi.co.jp
Mori
K
K
Yamaguchi
H
H
Kaniwa
H
H
Saisho
N
N
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
D009005:Monosaccharides; D011994:Recombinant Proteins; D005643:Fucose; D014269:Trifluoroacetic Acid; D010959:Tissue Plasminogen Activator; C085027:pamiteplase; D005944:Glucosamine; D008358:Mannose; D005690:Galactose