A systematic approach for reversed phase liquid chromatographic method development of fingolimod hydrochloride via design augmentation

Abstract

A stability indicating reversed phase liquid chromatographic (RP-LC) method was developed for the analysis of fingolimod hydrochloride (FIM) and its related compounds in drug substance and its dosage form. Separation of seven related compounds and three degradation products was achieved in a C18 stationary phase with 3 mM tetrabutyl ammonium hydrogen sulfate in 0.02% orthophosphoric acid and acetonitrile in gradient elution. Aqueous mobile phase components and column temperature were optimized through Design of experiments (DoE) for a fixed gradient program. Major degradation products were identified by LC-MS and degradation pathway is proposed. The validation results indicated that the method is sensitive in detection, linear, precise and accurate. The developed method was also demonstrated in UHPLC. Keywords: LC method development, Stability indicating, DoE, HPLC & UHPLC, Fingolimod and related substances