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-perfomance liquid chromatography (RP-HPLC) prior to analysis. Acid hydrolysis with TFA was performed at 100°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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