Abstract

NOVEL STABILITY INDICATING LIQUID CHROMATOGRAPHY METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF SIPONIMOD IN BULK SUBSTANCES AND PHARMACEUTICAL DOSAGE FORMS

Dr. Y. Narasimha Rao, Bala Hemanth* and Dr. M. Prasada Rao

ABSTRACT

Liquid chromatography is an analytical technique that is used to separate a certain sample into its individual components. HPLC is simple, specific, rapid, precise and accurate, it can be successfully and efficiently adopted for routine quality control analysis of drugs in bulk and pharmaceutical dosage form. In the present study a reverse phase high performance liquid chromatography method was developed and validated for the estimation Siponimod pharmaceutical formulations. To assess the effect of method parameters on chromatographic separation of the Siponimod, statistically designed experiments were performed by varying different method parameters such as buffer concentration, pH of mobile phase, flow rate, and column temperature. The separation was performed on ProntoSIL C18 column (250mm × 4.5 mm; 5μm) at room temperature using methanol, acetonitrile and phosphate buffer in the ratio of 65:25:10 (v/v)in isocratic condition at a flow rate of 1.0mL/min. The detection was performed by an ultraviolet detector (UVD) at 262nm with total run time of 10min. Calibration curves were linear in the concentration range of 5-30 μg/mL for with correlation coefficients of 0.9996. LOD and LOQ were found to be 0.02 μg/mL and 0.066 μg/mL proves the sensitivity of the developed method. The method can effectively separate the degradation compounds during the stress study and the standard drug Siponimod was found to be stable in all the stress degradation conditions. The developed method was able to determine the contents of the Siponimod commercial dosage forms and hence the method was used for the routine analysis of Siponimod in bulk drug as well as in pharmaceutical formulations.

Keywords: Siponimod, Selumetinib, Method validation, & High-Performance Liquid Chromatography.