Development of an enhanced uflc method for simultaneous determination of organic impurities and assay of amifampridine phosphate with greenness evaluation

The proposed research focuses on developing a simple, rapid and cost-effective chromatographic analytical method for the pharmaceutical evaluation of Amifampridine phosphate, a drug substance indicated for the symptomatic management of Lambert–Eaton myasthenic syndrome. The objective is to optimize a suitable analytical technique to enhance the multistage process control to obtain high pure drug substance by estimation of quantifying eight identified, unidentified impurities and assay. The analytical technique involves Heptafluoro butyric acid (HFBA), a MS compatible ion pair reagent buffer and methanol improves the retention of polar impurities using UFLC platform. The optimized chromatographic conditions applicable for simultaneous estimation of organic related impurities and assay using a Shim-pack Scepter C18-120 hybrid particle UFLC column, with 0.2% HFBA in water and methanol, operated under 40ºC column temperature at 260 nm in 18-minute gradient run. Its compatibility to LCMS helps to identify new impurities during process monitoring. The system suitability for impurities were set as per current pharmaceutical and regulatory guidelines, where the resolution and % RSD found > 1.5 and <5% respectively. Assay evaluation exhibited acceptable system suitability with a similarity factor between 0.98 and 1.02 and %RSD of 0.73. The Acceptable analytical greenness score obtained using AMGS, BAGI, AGREE and MoGAPI online tools. The reduction of analysis cost 78% for solvents with greater number of analyte over reported HPLC methods.