Quality by design (QbD) based development and validation of bioanalytical RP-HPLC method for dapagliflozln: Forced degradation and preclinical pharmacokinetic study

摘要

A rapid, sensitive, and reliable quality by design (QbD) based bioanalytical HPLC method was developed and validated for quantification of dapagliflozin (DFG) in rat plasma. The method was optimized using three levels and three-factor Box-Behnken statistical design (BBD). The mobile phase composition (acetonitrile % as A), flow rate (mL/min as B) and UV-wavelength (nm as C) selected as independent variable whereas area (AU as Y_1), retention time (min. as Y_2) and tailing factor (% as Y_3) selected as dependent variables. The developed method validated for linearity, sensitivity (LOD & LOQ), precision, accuracy, extraction recovery, matrix effect, and stability. Further the developed bioanalytical used to assess the pharmacokinetic study on a rat model. The developed method showed excellent linearity between 10 and 1200ng/mL with r~2 = 0.997, LOD (2.15ng/mL) and LOQ (6.52ng/mL). The validation results of the tested parameters were found within the acceptable limit. Further, DFG loaded S-SNEDDS showed the droplet size (75.34± 5.34 nm), PDI (0.426) and zeta-potential (-25.6 mV). The pharmacokinetic study results revealed the C_max of S-SNEDDS was 1.42 fold higher than DFG dispersion. The significantly high AUC value observed with S-SNEDDS indicated increased bioavailability of DFG as compared to DFG dispersion. From the study, it was concluded that QbD approach based bioanalytical method is suitable for the in-vitro and in-vivo estimation of DFG from bulk as well as developed formulations.