Development of a robust stability-indicating RP-HPLC method for simultaneous estimation of triple antidiabetic combination containing dapagliflozin, gliclazide and metformin

A straightforward, precise, accurate, and stability-indicating reverse-phase high-performance liquid chromatographic (RP-HPLC) approach was formulated and validated for the concurrent quantification of dapagliflozin, gliclazide, and metformin. Chromatographic separation was performed on a Kromasil C18 column (250 mm x 4.6 mm, 5 μm), utilizing a mobile phase of 20 mM ammonium acetate buffer (pH 3) and acetonitrile (35:65, v/v) under isocratic elution conditions. The analytes were identified with a UV detector at 224 nm. Metformin, dapagliflozin, and gliclazide were separated at 2.47 min, 3.90 min, and 6.46 min, respectively. The developed method was validated according to ICH Q2 (R2) guidelines. The method showed excellent linearity from 125 to 375 µg/mL for metformin, 2.5–7.5 µg/mL for dapagliflozin, and 15–45 µg/mL for gliclazide. The percentage recoveries for all three drugs were within acceptable limits, indicating the accuracy of the method. Precision studies revealed low relative standard deviation values, indicative of excellent repeatability and intermediate precision. Forced degradation studies were performed under acidic, alkaline, oxidative, thermal, and photolytic stress conditions to evaluate the stability-indicating capability of the method. The degradation products were adequately separated from the analyte peaks without interference, confirming the specificity and stability-indicating nature of the method. The proposed RP-HPLC method was found to be simple, rapid, reliable, and suitable for routine quality control analysis and stability studies of pharmaceutical formulations containing dapagliflozin, gliclazide, and metformin hydrochloride.