Development and Validation of Stability Indicating RP-HPLC Method for the Determination of Oseltamivir Phosphate in Oseltamivir Phosphate Capsules

Abstract

A simple, precise, accurate and stability-indicating HPLC method was developed and Validated for the determination of Oseltamivir Phosphate, which is used to treat Influenza in Pharmaceutical Dosage forms. The separation was achieved on YMC Pro C8, (150 x 4.6 mm) 5 µm column and potassium dihydrogen phosphate (pH 3.2±0.05) as a buffer,   Buffer: Methanol: Acetonitrile in the ratio 60:20:20 (v/v) as the Mobile Phase and Water, Methanol and acetonitrile 625:40:135 as a diluent. Chromatography was performed with sample injection volume of 20 µL, Flow rate 1.5mL/min and Column Temperature 45⁰C as Instrumental parameters and the analyte was detected at 207nm. The method has been validated for Specificity, Linearity, Accuracy, Precision, Solution Stability and Robustness. The Oseltamivir phosphate was subjected to stress conditions of hydrolysis (acid, base), oxidation (5% H₂O₂), thermal, humidity and photolytic degradations. The degradation was observed for Oseltamivir phosphate in Acidic, Alkaline and Oxidative Conditions, while, it is found stable to remaining stress condition. The calibration curve was found to be linear in the range of 76.6-229.7 μg/mL. The precision of the method was determined by inter-day and intraday variation studies having RSD values less than 1.0% showing high precision of the method. The validated RP-HPLC method was successfully applied to the quantitative determination of Oseltamivir Phosphate in capsule dosage form.