Abstract

SIMULTANEOUS ESTIMATION OF DAPAGLIFLOZIN AND SITAGLIPTIN BY RP-HPLC METHOD USING QUALITY BY DESIGN

Smita M. Mandlik*, L. A. Kawale and A. A. Wakchaure

Abstract

A new simple, sensitive, suitable, precise, and robust reversed phase high performance liquid chromatography (RP-HPLC) method was developed and validated for the simultaneous determination of sitagliptin and Dapagliflozin in it’s bulk and tablet dosage form using quality by design. Optimization of chromatographic condition was performed using central composite design. The separation was carried on HPLC system containing a UV- detector with Empower-3 software and Chemsil C18 column (250 mm x 4.6mm; 5μm). The detection was carried out at 243 nm. In the developed RP-HPLC method, Phosphate buffer : Acetonitrile : Methanol adjusted to a pH 5 (40:35:25 v/v/v) was used as mobile phase, at a flow rate of 1.0 mL/min and the retention time was 3.1 min and 10.0 min for Sitagliptin and Dapagliflozin respectively. The results of the analysis in the method were validated in terms of linearity, accuracy, precision, specificity, detection limit, quantification limit and robustness. The linearity of the developed method was confirmed over the concentration range of 10-50 μg/mL and 100-500 μg/mL with a correlation coefficient (r2) of 0.9989 and 0.9995 for Dapagliflozin and Sitagliptin respectively. The percentage recoveries of Sitagliptin and Dapagliflozin were found to be in the range of 98-100 % and 99-100 % respectively. The Detection limit and Quantitation limit of Sitagliptin and Dapagliflozin was found to be 16.45 μg/mL, 49.85 μg/mL and 2.03 μg/mL, 6.15 μg/mL respectively. The proposed method showed good predictability and robustness.

Keywords: RP-HPLC, QbD, Sitagliptin, Dapagliflozin, Method Development, Validation.