Abstract

ANALYTICAL METHOD DEVELOPMENT AND VERIFICATION OF DAPAGLIFLOZIN RELATED SUBSTANCES ESTIMATION IN DAPAGLIFLOZIN TABLETS BY RP-HPLC

Dr. Akuthota Ashok Kumar*, Bembadi Mukund Reddy, Bheemi Reddy Ashok Reddy and Dr. Rajesh Vooturi

ABSTRACT

The objective of the research is to develop a simple, sensitive, accurate, precise, and linear Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) Related Substances (RS) method for the quantification of Dapagliflozin related substances (degradants) and verify in Dapagliflozin tablets. The optimized method uses a reverse phase Agilent poroshell 120 EC- C18 column, (250 X 4.6 mm; 2.7μm), part number 690975-902, mobile phase-A consisting of 0.05% OPA buffer: Methanol in the proportion of 90:10 v/v, mobile phase-B consisting of water: acetonitrile in the proportion of 30:70 v/v, flow rate of 0.9 ml/min, injection volume of 5 µL, and detection wavelength of 225 nm. The developed method eluted Dapagliflozin at about 10 min by gradient elution mode. Dapagliflozin (unknown impurity) exhibited linearity in the range LOQ of 0.05% (0.25 μg/ml) to 150% (1.6μg/ml) considering the unknown impurity specification limit as 0.2%. The precision is exemplified by relative standard deviation of 2.3% and 2.5% for single maximum known impurity and total impurities. Percentage individual recovery was found to be in the range of 98.86 and 101.96 during accuracy studies. Specificity established non-interference of blank, placebo, known impurities with the degradants and Dapagliflozin peak along with the resolution between the known impurities and the degradants. Peak purity passed for Dapagliflozin peak in diluted standard, as such sample, spiked sample and forced degradation sample, indicating that method developed is a stability-indicating and hence this method can be explored for the routine analysis of Dapagliflozin related substances quantification in Dapagliflozin tablets in various pharmaceutical industries.

Keywords: HPLC, Dapagliflozin Tablets, Related substances, Impurities, Degradants.