Validation of an RP-LC Method for the Determination of Interferon-2a in Pharmaceutical Formulations

A reversed phase liquid chromatography (RP-LC) method was validated for the determination of interferon-2a in pharmaceutical formulations. The RP-LC method was carried out on a Jupiter C4 column (250mm4.6mm I.D.), maintained at room temperature. The mobile phase A consisted of 0.1% TFA and the mobile phase B was acetonitrile with 0.1% TFA, run at a flow rate of 1mL/min, and using photodiode array (PDA) detection at 214nm. The chromatographic separation was obtained with the retention time of 32.6min, and was linear in the concentration range of 0.5-50 MIU/mL (r2=0.9999). The specificity was proven through degradation studies, which also showed that there was no interference of the excipients. The accuracy was 100.84% with bias lower than 1.87%. The limits of detection and quantitation were 0.19 and 0.5 MIU/mL, respectively. Moreover, method validation demonstrated acceptable results for precision and robustness. The proposed method was applied for the analysis of the interferon-2a and their related proteins in parenteral dosage forms, contributing to establishing alternatives to improve the quality control assuring the therapeutic efficacy of the biological medicine.