SIMULTANEOUS ESTIMATION OF AMANTADINE HYDROCHLORIDE AND OSELTAMIVIR PHOSPHATE USING PRECOLUMN DERIVATIZATION TECHNIQUE

A simple, efficient HPLC method has been developed and validated for simultaneous determination of Amantadine hydrochloride and Oseltamivir phosphate using pre-column derivatization with phenyl isothiocyanate (PITC). The chromatography separation was achieved on Phenomenex C18 column of 250 × 4.6 mm 5 µm particle size, mobile phase used for chromatographic run consisted of acetonitrile and water (60: 40% v/v) with a flow rate of 2 mL/min. The analysis was performed at ambient temperature, and the eluent was monitored at 250 nm using a UV detector. The retention time of Amantadine hydrochloride and Oseltamivir phosphate was found to be 7.1 min and 3.2 min. The limit of detection (LOD) for Amantadine hydrochloride and Oseltamivir phosphate was observed to be 0.1µg/mL and 0.25 µg/mL, respectively. Limit of quantification (LOQ) was found to be 0.5 µg/mL and 0.25 µg/mL. The developed method was validated as per ICH guidelines using parameters like linearity, specificity, precision, linearity, accuracy, ruggedness, robustness, LOD, and LOQ. All the validation parameters were found to be well within the acceptance criteria. Hence, the proposed method can be used for the routine analysis of Amantadine hydrochloride and Oseltamivir phosphate in bulk and tablet dosage forms.