A generic novel HPLC method for the determination of nitrite ion by direct derivatization

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BackgroundThis research introduces a novel, simplified method to quantify carcinogenicity generating nitrite in Key Starting Materials (KSMs) and specific drugs. Since no straightforward approach exists in the literature, this method aimed to fill the lacunae and fulfill safety assessments in pharmaceutical applications. MethodThe developed method employs a direct and simple derivatization procedure using High-Performance Liquid Chromatography (HPLC). The conditions include Zorbax SB C8 column (stationary phase), trifluoroacetic acid (TFA): water: acetonitrile at 0.05:55:45 % v/v as mobile phase A, and TFA: acetonitrile at 0.05:100 % v/v as mobile phase B in a gradient elution at 0.8 mL/minute flow rate. The gradient program is time/%A, 0/100, 10/100, 15/0, 19/0, 20/100, 25/100. Significant findingsA simple method was established to regulate the nitrite ion levels in KSMs and certain medications. By making a fine-tuning to chromatographic conditions, this method can quantify nitrite content in any pharmaceutical materials, thus, demonstrating its generic applicability. The developed method was validated according to the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines. It is linear and accurate from the limit of quantitation (LOQ) to 150 % (i.e., 0.5 to 1.5 ppm) of the specified limit (1.0 ppm). The LOQ and limit of detection (LOD) for this method are 0.5 and 0.1 ppm, respectively.