Design of Experiment-Based Rp-Hplc Bioanalytical Method Development and Validation for the Estimation of Linagliptin and Dapagliflozin in Bulk and in Their Combined Dosage Form

Resumen

The aim of this study was to develop and optimize a simple, cost-effective, and robust Bioanalytical RP-HPLC method by Design of Experiment based Box–Behnken design for the simultaneous estimation of Linagliptin and Dapagliflozin in human plasma. The optimal chromatographic separation was achieved having C18 (Thermo) column (250 mm × 4.6 mm, 5 μ) and using mobile phase as Methanol and 0.1 % OPA (80:20) with a flow rate 0.9 ml/min and UV detection at 242 nm. A Box-Behnken design was used to test robustness of the method with describes the interrelationship of Mobile phase, Flow rate and Wavelength at three different levels. The method was found to be linear in the range of 2-10 μg/mL (R² >0.9997) and 4-20 μg/mL (R² >0.9994) for Linagliptin and Dapagliflozin, respectively. The developed bioanalytical method was validated as per recommended ICH guidelines which revealed the high degree of linear, precise, accurate, sensitive and robust method over the existing RP-HPLC method for Vildagliptin and Pioglitazone hydrochloride.