Development and Validation of Oseltamivir Carboxylate

Abstract

Objective: To develop a simple HPLC analysis for the quantification of oseltamivir carboxylate (OC) and  oseltamivir phosphate (OP) in human plasma.  Method: Chromatographic separation was carried out using Hypersil BDS cyano 5 μm column (length 250 mm, inner diameter 4.6 mm) with UV detection at 230 nm. The gradient system composed of 50 mM ammonium acetate: acetonitrile (95:5) for 4 min, followed by 50 mM ammonium acetate: acetonitrile (70:30) for 7 min, and finally 50 mM ammonium acetate: acetonitrile (95:5) for 4 min.  Results: Coefficients of determination for the quantification of OC and OP were 0.9994 and 0.9991, respectively, with retention times of 4.35 and 11.04 min, respectively. The method limit of quantification for both compounds was 1.5  μg/ml. Coefficients of variation for the between- and within-day precision of OC were 2.15 - 7.19% and 1.97 - 6.63%, respectively, and those of OP were 2.41 - 8.52% and 1.90 - 8.57%, respectively. Conclusion: A simple and reliable HPLC quantification method of oseltamivir carboxylate and oseltamivir phosphate in human plasma was successfully validated. Keywords: oseltamivir carboxylate, oseltamivir phosphate, HPLC, method validation