Stability Indicating Simultaneous Method development and Validation of Dapagliflozin and Saxagliptin by RP-HPLC

Headnote

ABSTRACT:

The developed method was validated according to ICH guidelines with respect to specificity, linearity, limits of detection, quantification, accuracy, precision, and robustness. The stability indicating simultaneous method development and validation of Dapagliflozin and Saxagliptin by RP-HPLC in drug substance and product. The chromatographic separation was done in an isocratic mode using the BDS C18 (150 x 4.6mm, 5.0µ) column with mobile phase containing a Ammonium acetate buffer: ACN (40:60 %v/v) used for efficient chromatographic separation. The flow rate of the mobile phase was adjusted to 1.0mL/min with 30°C column temperature and detection of wavelength at 220nm, injection volume 10µl was fixed for achieving good elution of eluents. The retention time for DAPA and SAXA were found to be 2.207 min and 2.889 min respectively. DAPA and SAXA were linear in the concentration ranges from 0-15µg/ml and 0 -8µg/ml respectively. Regression analysis showed that R² value (correlation coefficient) for DAPA and SAXA were found to be 0.999. LOD and LOQ of DAPA was found to be 21.6µg/ml and 9.1µg/ml, SAXA was found to be 11.5µg/ml 9.1µg/ml. The developed method was validated and found to be accurate, specific and robust. Both the drugs were subjected to the stress conditions like acidic, basic, oxidative, photolytic and thermal conditions. The degradation results were found to be satisfactory. In acidic, alkaline and peroxide stress conditions. The forced degradation test results confirmed that the DAPA and SAXA peaks were homogenous in all stress conditions, thus proving the stability-indicating nature of the method. This method could be applied for the simultaneous estimation of DAPA and SAXA in drug substance.

KEYWORDS: RP-HPLC, Dapagliflozin (DAPA) and Saxagliptin (SAXA), Stability indicating, Analytical method development, Analytical method validation.

INTRODUCTION:(1-3)

Dapagliflozin (DAPA), 2S,3R,4R,5S,6R)-2-(4-Chloro3-(4-ethoxybenzyl) phenyl)-6- (hydroxymethyl) tetrahydro-2H- pyran-3,4,5-trio is widely used for the treatment of type 2 diabetes mellitus, It shows its action by reducing glucose levels by excreting through kidney. Dapagliflozin molecular formula is C21H25ClO6, and molecular weight is 408.873gram/mole.

Saxagliptin (SAXA) is competitive dipeptidyl peptidase-4 inhibitor used to treat diabetes mellitus type2. Its empirical formula is C18?5N3O2,and corresponding molecular weight of the compound 315.41gram/mole.

The combination of DAPA and SAXA is used to treat diabetes mellitus type-2, The chemical structures of DAPA and SAXA and are shown in Image 1.

DAPA and SAXA...