A simple isocratic RP-HPLC method for quality control of oseltamivir capsules

Agim Ameti, Jasmina Slavkovska, Katerina Starkoska, Zorica Arsova-Sarafinovska


A simple isocratic reversed-phase high performance liquid chromatographic (RP-HPLC) method was developed for determination of oseltamivir active pharmaceutical ingredient (API) in bulk drug and pharmaceuticals. The separation was achieved on a Purospher STAR® RP – 18e column with a mobile phase consisting of methanol- 0.02 mol l-1 phosphate buffer, pH 5, 50:50 (v/v). Chromatographic results demonstrated the specificity of the method for determination of oseltamivir in presence of degradation products generated in studies of forced decomposition. The limit of detection (LOD) and limit of quantification (LOQ) for oseltamivir phosphate were 0,0162 μg ml-1 and 0,0491 μg ml-1, respectively. The advantages of this method include simple sample treatment and short elution time (less than 6 min). Furthermore, using methanol instead of acetonitrile in a mobile phase composition considerably reduces the laboratory expenses, still retaining adequate sensitivity for routine analysis as well as for evaluation of potentially counterfeit Tamiflu® products.



Oseltamivir; RP-HPLC; HPLC-DAD; Quality control; Pharmaceuticals

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Writing Committee of the WHO Consultation on Clinical Aspects of Pandemic (H1N1) 2009 Influenza. Clinical Aspects of Pandemic 2009 Influenza A (H1N1) Virus Infection. N. Engl. J. Med. 2010, 362, 1708–1719.

Novel Swine-Origin Influenza A (H1N1) Virus Investigation Team. Emergence of a novel swine origin influenza A (H1N1) virus in humans. N. Engl. J. Med. 2009, 360, 2605-2615.

Y. Itoh, K. Shinya, M. Kiso, et al. In vitro and in vivo characterization of new swine origin H1N1 influenza viruses, Nature 460, 1021-1025 (2009).

M.L. Lim, C.Y. Chong, W.S. Tee, W.Y. Lim, J.J. Chee. Influenza A/H1N1 (2009) infection in pregnancy — an Asian perspective. BJOG 117, 551-556 (2010).

WHO guidelines for pharmacological management of pandemic (H1N1) 2009 influenza and other influenza viruses. Geneva: World Health Organization, February 2010. http://www.who.int/csr/resources/publications/swineflu/h1n1_use_antivirals_20090820/en/index.html. (07 March 2012, date last accessed).

Tamiflu SmPC. Tamiflu Summary of Product Characteristics. http://emc.medicines.org.uk (7 March 2012, date last accessed)

B.E. Davies. Pharmacokinetics of oseltamivir: an oral antiviral for the treatment and prophylaxis of influenza in diverse populations, J. Antimicrob. Chemother. 65, 5–10 (2010).

M.E. Bosch, C. Bosch Ojeda, A.J. Ruiz Sánchez, F. Sánchez Rojas. Analytical methodologies for the determination of oseltamivir, Res. J. Pharm. Biol. Chem. Sci. 1 (3), 368-376 (2010).

M.J. Navas, A.M. Jimenez. Analytical methods to determine anti-influenza drugs, Crit. Rev. Anal. Chem. 41, 81-97 (2011).

H. Wiltshire, B. Wiltshire, A. Citron, A. T. Clarke, C. Serpe, D. Gray, et al. Development of a high-performance liquid chromatographic mass spectrometric assay for the specific and sensitive quantification of Ro 64–0802, an anti-influenza drug, and its pro-drug, oseltamivir, in human and animal plasma and urine. J. Chromatogr. B Biomed. Sci. Appl. 745, 373–388 (2000).

N. Lindegardh, T.T. Hien, J. Farrar, P. Singhasivanon, N.J. White, N.P. Day. A simple and rapid liquid chromatographic assay for the evaluation of potentially counterfeit Tamiflu®, J. Pharm. Biomed. Anal. 42, 430–433 (2006).

J. Joseph-Charles, C. Geneste, E. Laborde-Kummer, R. Gheyouche, H. Boudis, J.P. Dubost. Development and validation of a rapid HPLC method for the determination of oseltamivir phosphate in Tamiflu and generic versions. J. Pharm. Biomed. Anal. 44, 1008-1013 (2007).

C. Fuke, Y. Ihama, T. Miyazaki. Analysis of oseltamivir active metabolite, oseltamivir carboxylate, in biological materials by HPLC-UV in a case of death following ingestion of Tamiflu, Leg. Med. 10, 83-87 (2008).

G. Bahrami, B. Mohammadi, A. Kiani. Determination of oseltamivir carboxylic acid in human serum by solid phase extraction and high performance liquid chromatography with UV detection, J. Chrom. B: Anal. Technol. Biomed. Life Sci. 864, 38-42 (2008).

B. Narasimhan, K. Abida, K. Srinivas. Stability indicating RP-HPLC method development and validation for oseltamivir API, Chem. Pharm. Bull. (Tokyo) 56, 413-417 (2008).

Z. Aydogmus, S. Caglar, S. Toker. RP-HPLC method for determination of oseltamivir phosphate in capsules and spiked plasma, Anal. Lett. 43, 2200-2209 (2010).

ICH Q2R1: Validation of Analytical Procedures: Text and Methodology. Proceeding of the International Conference on Harmonization of Technical Requirements for the Registration of Drugs For Human Use, Geneva, Switzerland, 1996.

ICH, Guideline on Analytical Method Validation, Proceeding of International Convention on Quality for the Pharmaceutical Industry, Toronto, Canada, 2002.

General Chapter <1225>, Validation of compendial methods, United States Pharmacopeia 32, National Formulary 27, Rockville, Md., USA, The United States Pharmacopeial Convention, Inc., (2009).

General Chapter <621>, Chromatography, United States Pharmacopeia 32, National Formulary 27, Rockville, Md., USA, The United States Pharmacopeial Convention, Inc., (2009).

Center for Drug Evaluation and Research. Food and Drug Administration. Reviewer Guidance: Validation of Chromatographic Methods. Washington, DC: Department of Health and Human Services; 1994.

ICH Q1A: Stability Testing of New Drug Substances and Products. (International Conference on Harmonization of Technical Requirements for the Registration of Drugs For Human Use, Geneva, Switzerland, February 2003).

DOI: http://dx.doi.org/10.20450/mjcce.2012.10


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