Validated Stability Indicating RP-HPLC for Quantitation of Nitazoxanide in Presence of Its Alkaline Degradation Products and Their Characterization by HPLC-Tandem Mass Spectrometry

A simple and sensitive stability indicating HPLC method was devel-oped and validated for quantitative determination of Nitazoxanide (NTZ), a new antiprotozoal drug, in presence of degradation products generated under forced alkaline hydrolysis. Chromatographic separ-ation was achieved on Inertsil C8-3 column (1503 4.6 mm i.d.) using a mobile phase composed of acetonitrile: 50 mM ammonium acetate buffer (50:50, v/v, pH 5.0 adjusted with acetic acid) at a flow rate of 1 mL/min. Quantification was achieved with UV detection at 298 nm based on relative peak area. The method was linear over the concen-tration range of 0.8–50 mg/mL (r5 0.9999) with a limit of detection and quantification 0.0410 and 0.1242 mg/mL, respectively. The devel-oped method has the requisite accuracy, selectivity, sensitivity and precision to assay NTZ in presence of its degradation products either in bulk powder or in pharmaceutical formulations. The degradation products were then identified by HPLC-MS/MS analysis using an elec-trospray ionization source and an ion trap analyzer