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Talanta ; 94: 1-7, 2012 May 30.
Article de Anglais | MEDLINE | ID: mdl-22608407

RÉSUMÉ

A stability-indicating capillary zone electrophoresis (CZE) method was validated for the analysis of granulocyte-macrophage colony-stimulating factor (rhGM-CSF) using leuprorelin acetate (LA), as internal standard (IS). A fused-silica capillary (75 µm i.d.; effective length, 72 cm) was used at 25 °C; the applied voltage was 12 kV. The background electrolyte solution consisted of 50mM di-sodium hydrogen phosphate solution at pH 8.8. Injections were performed using a pressure mode at 50 mbar for 9s, with detection by photodiode array detector set at 200 nm. Specificity and stability-indicating capability were established in degradation studies, which also showed that there was no interference of the excipients. The method was linear over the concentration range of 2.5-200 µg mL(-1) (r(2)=0.9995) and the limit of detection (LOD) and limit of quantitation (LOQ) were 0.79 µg mL(-1) and 2.5 µg mL(-1), respectively. The accuracy was 99.14% with bias lower than 1.40%. The method was applied to the quantitative analysis of biopharmaceutical formulations, and the results were correlated to those of a validated reversed-phase LC method (RP-LC), and an in vitro bioassay, showing non-significant differences (p>0.05).


Sujet(s)
Électrophorèse capillaire/méthodes , Facteur de stimulation des colonies de granulocytes et de macrophages/analyse , Protéines recombinantes/analyse , Dosage biologique , Calibrage , Lignée cellulaire , Chromatographie en phase inverse , Humains , Concentration en ions d'hydrogène , Leuprolide/analyse , Limite de détection , Phosphates/composition chimique , Silice/composition chimique
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