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

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• 作者：Sergio Luiz Dalmora ; ^{sdalmora@terra.com.br} ; Cairo dos Santos Butzge ; Francine Trevisan Machado ; Maurí ; cio Elesbã ; o Walter ; Maria Elisabeth de Á ; vila Dalmora ; Ricardo Bizogne Souto
• 关键词：Capillary zone electrophoresis ; Reversed-phase liquid chromatography ; Recombinant human granulocyte-macrophage colony-stimulating factor ; Cell culture ; Biotechnology
• 刊名：Talanta
• 出版年：2012
• 期刊代码：106_00399140
• 类别：ch
• 出版时间：30 May, 2012
• 卷：94
• 期：Complete
• 页码：1-7
• 文件大小：418 K

摘要

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 渭m i.d.; effective length, 72 cm) was used at 25 掳C; the applied voltage was 12 kV. The background electrolyte solution consisted of 50 mM di-sodium hydrogen phosphate solution at pH 8.8. Injections were performed using a pressure mode at 50 mbar for 9 s, 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 渭g mL^鈭? (r² = 0.9995) and the limit of detection (LOD) and limit of quantitation (LOQ) were 0.79 渭g mL^鈭? and 2.5 渭g mL^鈭?, 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).