Department of Industrial Pharmacy, Federal University of Santa Maria, 97105-900 Santa Maria, RS, Brazil.
Talanta. 2012 May 30;94:1-7. doi: 10.1016/j.talanta.2012.03.015. Epub 2012 Mar 11.
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).
一种用于分析粒细胞-巨噬细胞集落刺激因子（rhGM-CSF）的稳定性指示毛细管区带电泳（CZE）方法已经过验证，使用醋酸亮丙瑞林（LA）作为内标（IS）。使用熔融石英毛细管（75μm id；有效长度，72cm），在 25°C 下；施加电压为 12 kV。背景电解质溶液由 50mM 二磷酸氢二钠溶液组成，pH 值为 8.8。采用压力模式在 50 mbar 下进样 9s，用光电二极管阵列检测器在 200nm 处检测。在降解研究中确定了特异性和稳定性指示能力，这也表明赋形剂没有干扰。该方法在 2.5-200μgmL（-1）（r（2）=0.9995）浓度范围内呈线性，检测限（LOD）和定量限（LOQ）分别为 0.79μgmL（-1）和 2.5μgmL（-1）。准确度为 99.14%，偏差低于 1.40%。该方法已应用于生物制药制剂的定量分析，结果与经过验证的反相液相色谱法（RP-LC）和体外生物测定法相关联，显示无显著性差异（p>0.05）。
