title: Stability-indicating capillary zone electrophoresis method for the assessment of recombinant human granulocyte-macrophage colony-stimulating factor and its correlation with reversed-phase liquid chromatography method and bioassay.
abstract: 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).
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil. sdalmora@terra.com.br
authors.forename: Sergio Luiz
authors.initials: SL
authors.lastname: Dalmora
authors.forename: Cairo dos Santos
authors.initials: Cdos S
authors.lastname: Butzge
authors.forename: Francine Trevisan
authors.initials: FT
authors.lastname: Machado
authors.forename: Maurício Elesbão
authors.initials: ME
authors.lastname: Walter
authors.forename: Maria Elisabeth de Ávila
authors.initials: ME
authors.lastname: Dalmora
authors.forename: Ricardo Bizogne
authors.initials: RB
authors.lastname: Souto
mesh_terms: D001681:Biological Assay; D002138:Calibration; D002460:Cell Line; D056148:Chromatography, Reverse-Phase; D019075:Electrophoresis, Capillary / Q000379:methods*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016729:Leuprolide / Q000032:analysis; D057230:Limit of Detection; D010710:Phosphates / Q000737:chemistry; D011994:Recombinant Proteins / Q000032:analysis*; D012822:Silicon Dioxide / Q000737:chemistry
chemical_list: D010710:Phosphates; D011994:Recombinant Proteins; D012822:Silicon Dioxide; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D016729:Leuprolide; C018279:sodium phosphate