Stability Indicating RP-HPLC Method Development and Validation for Oseltamivir API

Abstract

Oseltamivir is an antiviral drug used in the treatment and prophylaxis of influenza virus A and B infections. It was developed by Gilead Sciences and is currently marketed by Haffmann-La Roche (Roche) under the trade name Tamiflu. Oseltamivir is readily absorbed from the gastrointestinal tract after oral administration of oseltamivir phosphate. It is extensively converted to oseltamivir carboxylate by esterases located predominantly in the liver. Oseltamivir carboxylate is eliminated entirely by renal excretion. Less than 20% of an oral radiolabeled dose is eliminated in feces.1) Chemically it is (3R,4R,5S)-4-acetylamino-5-amino-3-(1-ethylpropoxy)-1-cyclohexene-1-carboxylic acid, ethyl ester. 2)Stability testing forms an important part of the process of drug product development. The purpose of stability testing is to provide evidence on how the quality of a drug substance varies with time under the influence of a variety of environmental factors such as temperature, humidity and light, and enables recommendation of storage conditions, retest periods, and shelf life to be established. The assay of oseltamivir API (Active Pharmaceutical Ingredient) in stability test sample needs to be determined using stability indicating method, as recommended by the International Conference on Harmonization (ICH) guidelines 3) and USP.4)The present work was designed to develop a simple, precise and rapid analytical LC procedure, which would serve as stability indicating assay method for analysis of oseltamivir API. Literature survey for oseltamivir analysis revealed several methods based on different techniques, viz. LC-MS assay for it's quantification in plasma and urine, 5) LC assay for evaluation of Tamiflu 6) and HPLC method for the determination of oseltamivir in Tamiflu. 7)The present study is the first time report on stability indicating assay of oseltamivir in presence of degradation products by HPLC. In this method gradient elution method is selected for the analysis of oseltamivir API because it gave better base line separation and peak width, which is suitable for the routine analysis of oseltamivir. Although the gradient elution method uses two-pump system, the gradient run provided a proper peak with baseline, which prompted us to select this method for the analysis of oseltamivir API.In view of above in the present study we hereby report the development and validation of a stability indicating gradient reverse-phase HPLC (RP-HPLC) method for analysis of oseltamivir API in presence of degradation products as per ICH guidelines.In order to establish the stability indicating nature of the method, force degradation of oseltamivir was performed under various stress conditions (basic, acidic and oxidative), and stressed samples were analyzed by the proposed method. The proposed LC method was able to separate the drug from degradation products generated during forced degradation studies. The developed method was validated as per ICH guidelines 8) and its updated international convention.9) The linearity of response, preci...

Determinação rápida de oseltamivir em cápsulas por cromatografia líquida de ultraeficiência com detector por arranjo de diodos

“…Comparado aos métodos descritos na literatura com emprego da CLAE ocorreu redução de 4 vezes, pelo menos, no tempo de análise cromatográfica. 1,2,8,[10][11][12] No presente trabalho o tempo despendido na preparação das amostras foi de aproximadamente 40 min, considerando-se os tempos de incubação no ultrassom e centrifugação, de 10 min cada. No trabalho apresentado por Lindegardh et al 10 para a determinação de OSV em especialidades farmacêuticas, empregando CLAE com detecção por UV, o tempo para a preparação das amostras (de cerca de 60 min) e de corrida cromatográfica (de 7 min) foram superiores aos propostos por este trabalho.…”

“…No trabalho apresentado por Lindegardh et al 10 para a determinação de OSV em especialidades farmacêuticas, empregando CLAE com detecção por UV, o tempo para a preparação das amostras (de cerca de 60 min) e de corrida cromatográfica (de 7 min) foram superiores aos propostos por este trabalho. 10 Nos métodos desenvolvidos por Narasimhan et al 2 e JosephCharles et al, 11 que também utilizaram CLAE-UV para avaliar cápsulas de similares de OSV, apesar do preparo simples e rápido das amostras, as corridas cromatográficas foram de 12 e 5 min, com vazão da fase móvel de 1 e 1,2 mL/min, respectivamente. Por fim, Green et al 8 utilizaram colorimetria seguida por CLAE para avaliação das cápsulas de produtos genéricos de OSV, sendo que a CLAE teve tempo de análise cromatográfica de 7 min, com vazão de fase móvel de 1 mL/min.…”

“…2,3 Segundo a Organização Mundial da Saúde (OMS), a dose recomendada para o tratamento de adultos é de 75 mg duas vezes ao dia, por um período de 5 dias, e, para a profilaxia é de 75 mg uma vez ao dia, por pelo menos 7 dias.…”

Analysis of Oseltamivir in Tamiflu Capsules Using Micellar Electrokinetic Chromatography

“…3) Bioassay, 4) chromatographic methods with fluorescence, 5) mass spectrometric 6,7) and UV [8][9][10][11][12][13] detection have been described for the analysis of oseltamivir, but to the best of our knowledge, there is no report on the capillary electrophoresis (CE) method for the analysis of oseltamivir. To provide more information on the reported methods to analyze oseltamivir and its hydrolytic product, the related articles are briefly reviewed in chronological order.…”

“…The reported LOD and LOQ were 0.05 and 0.3 ng/ml, respectively. 13) The main advantages of CE methods are speed, resolution, efficiency, analyte solubility and stability, minimal reagent and solvent consumption, compatibility with mass spectrometry, and the availability of several modes which have all made CE a popular analytical technique in the field of pharmaceutical and biomedical analysis. The applications of various aspects of CE in analysis of small-molecule pharmaceuticals were reviewed by Altria and coworkers.…”