Development and Validation of a Stability-Indicating Size-Indicating Size-Exclusion LC Method for the Determination of rhIFN-α2a in Pharmaceutical Formulations

Abstract

A size-exclusion LC method was validated for the determination of interferon-a2a (rhlFN-alpha2a) in pharmaceutical formulations without interference from human serum albumin. Chromatographic separation was performed on a BioSep-SEC-S 2000 column (300 x 7.8 mm id). The mobile phase consisted of 0.001 M monobasic potassium phosphate, 0.008 M sodium phosphate dibasic; 0.2 M sodium chloride buffer, pH 7.4, run at a gradient flow rate and using photodiode array detection at 214 nm, was used. Chromatographic separation was achieved with a retention time of 17.2 min, and the analysis was linear over the concentration range of 1.98 to 198 microg/mL (r2 = 0.9996). The accuracy was 101.39%, with bias lower than 1.67%. The LOD and LOQ were 0.87 and 1.98 microg/mL, respectively. Moreover, method validation demonstrated acceptable results for precision and robustness. The method was applied to the assessment of rhlFN-alpha2a and related proteins in biopharmaceutical dosage forms, and the content/potencies were correlated to those given by a validated RP-LC method and an in vitro bioassay. It was concluded that use of the methods in conjunction allows a great improvement in monitoring stability and QC, thereby ensuring the therapeutic efficacy of the biotechnology-derived medicine.

Desenvolvimento de processo de produção e caracterização de interferon-&alpha;2a secretado no espaço periplásmico de <i>Escherichia Coli</i>

“…São descritas na literatura diversas técnicas de análise de IFN a partir de amostras puras, preparações farmacêuticas, e até mesmo de amostras de soro (CUETO-ROJAS et al, 2010;EL GHAZALY et al, 2013;SILVA et al, 2009;ZARRIN et al, 2006;ZIMMERMANN et al, 2013…”

“…Muitos autores sugerem o uso de técnicas cromatográficas para avaliar concentração, estabilidade e pureza de IFNα-2a e IFNα-2b a partir de preparações farmacêuticas Zimmermann et al (2013). validaram um método de HPLC de exclusão molecular para determinação de IFNα-2a em formulações farmacêuticas.…”