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  • Title: Monosaccharide composition analysis of pamiteplase by anion exchange chromatography with pulsed amperometric detection.
    Author: Yokota H, Mori K, Yamaguchi H, Kaniwa H, Saisho N.
    Journal: J Pharm Biomed Anal; 1999 Dec; 21(4):767-74. PubMed ID: 10701942.
    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.
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