A Pre-Column Derivatization Technique for The Development and Validation of a HPLC-UV Method for the Determination of Amantadine Hydrochloride in Bulk and Formulations by using (2-Napthoxy) Acetyl Chloride

Author(s): Geetharam Yanamadala, N. Lavanya and P. Praveen Srikumar

A simple, precise, and accurate HPLC method has been developed and validated for the quantitative analysis of amantadine hydrochloride in tablet form by using (2-Napthoxy) Acetyl chloride as derivatization agent and memantine as an internal standard. An isocratic separation was achieved using inertsil ODS-3V, 250 x 4.6, 5 μm column with a flow rate of 1.5 mL/min and UV detector at 226 nm. The mobile phase consisted of 0.02 M ammonium acetate buffer and methanol in the ratio (12:88). The retention time for Amantadine and Memantine was around 6.23 and 8.62. The method was validated for specificity, linearity, precision, accuracy, robustness, and solution stability. The specificity of the method was determined by assessing interference from the placebo. The method was linear over the concentration range 28-171 μg/mL (r2 = 0.999) with a Limit of Detection (LOD) and Limit of Quantitation (LOQ) 0.23 and 0.69 μg/mL, respectively. The accuracy of the method was between 98.9-99.6%. The method was found to be robust and suitable for the quantitative analysis of amantadine hydrochloride in a tablet formulation

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