Analysis of Insulin Glargine in Biopharmaceutical Formulations by Validated RP-LC and SE-LC Methods

作者：Schramm Vanessa G; Xavier Bruna; Walter Mauricio E; Perlin Valquiria G; Motta Luis G J; Cardoso Leonardo C; Dalmora Sergio L

来源：Latin American Journal of Pharmacy, 2017, 36(2): 332-339.

• biotechnology
• insulin glargine
• reversed-phase liquid chromatography
• size exclusion liquid chromatography
• validation

A reversed-phase liquid chromatography (RP-LC) and a size exclusion liquid chromatography (SE-LC) methods were validated for the assessment of insulin glargine. The RP-LC method was carried out on a Jupiter-C-4 column (250 mm x 4.6 mm i.d.), at 30 degrees C. The mobile phase consisted of a 50 mM sodium sulphate buffer solution pH 2.5, and acetonitrile, run isocratically at a flow rate of 0.5 mL/min. The SE-LC method was carried out on a BioSep-SEC-S 2000 column (300 x 7.8 mm i.d.), at 25 degrees C. The mobile phase consisted of 30 mM MES buffer pH 2.5, run isocratically at a flow rate of 0.6 mL/min. The separation was obtained with retention times of 7.5 and 9.9 min, with quantitation limits of 0.054 and 0.027 mu g/mL, and detection at 214 and 200 nm, respectively, for RP-LC and SE-LC. The methods were applied to the analysis of biotherapeutics.

• 出版日期2017