QbD-Driven Analytical Method Development and Validation for Linagliptin and Empagliflozin by RP-HPLC

Abstract

The current investigation focused on the development and validation of an analytical method driven by Quality by Design (QbD) for the simultaneous analysis of Linagliptin and Empagliflozin utilizing RP-HPLC. A 2FI factorial design was utilized to identify and optimize key method variables, specifically the composition of the mobile phase (% methanol) and the flow rate. Methanol and acetonitrile were used as the mobile phase in an isocratic chromatographic separation process on a C18 column. For both medications, the wavelength of detection was set at 289 nm. Specificity, linearity, precision, accuracy, robustness, LOD, and LOQ were among the parameters that were examined during the validation process in accordance with the ICH Q2(R1) guidelines. The developed technique exhibited distinct, well-defined peaks for Linagliptin and Empagliflozin, with retention times within acceptable parameters. Linearity was confirmed across a concentration range of 20-80ug/ml for Empagliflozin and Linagliptin analysis, with correlation coefficients (R²) of 0.999 for both compounds, demonstrating an outstanding linear response. The method showed high accuracy, with recovery rates ranging from 98–102%, and precision with %RSD values below 2%. The LOD and LOQ values verified the method’s sensitivity. ANOVA analysis indicated that the mobile phase and flow rate significantly impacted retention time, theoretical plates, and tailing factors. No interference was detected in blank chromatograms, ensuring specificity. The method proved robust against minor deliberate variations in chromatographic conditions. The research effectively developed a QbD-based RP-HPLC technique that is straightforward, accurate, precise, and dependable for the concurrent estimation of Linagliptin and Empagliflozin. This method is appropriate for regular quality control assessments and stability investigations of pharmaceutical products.