Objective: The study aimed to develop and validate a stability-indicating RP-HPLC method for the estimation of related substances in Leuprolide acetate depot formulations using an Analytical Quality by Design (AQbD) approach. Materials and Methods: Critical method parameters such as flow rate, column oven temperature, and mobile phase pH were screened and optimized using a Central Composite Design. Preliminary method development included risk assessment to identify critical analytical attributes. The optimized method was validated as per ICH guidelines for specificity, linearity, accuracy, precision, robustness, and stability. Results and Discussion: Optimization studies revealed that flow rate, temperature, and pH significantly influenced resolution and peak tailing. The method demonstrated acceptable resolution and peak symmetry, with satisfactory ANOVA values confirming the suitability of the design model. Validation results established the method as specific, precise, linear, accurate, robust, and stable, fulfilling ICH acceptance criteria. Conclusion: The developed RP-HPLC method, guided by AQbD principles, proved to be reliable and stability-indicating for quantification of related substances in Leuprolide acetate depot formulations. This method offers a robust tool for routine quality control and stability studies in pharmaceutical analysis..
Keywords: Leuprolide acetate; Analytical Quality by Design (AQbD); RP-HPLC; Related substances; Stability-indicating method; Method validation
How to cite this article:Mandale VR, Dalvi SD, Tekade AR, Analytical Qbd Approach For Development And Validation Of Stability-Indicating Rp-Hplc Method For Related Substances In Leuprolide Acetate Depot Formulations.Int J Drug Deliv Technol. 2026;16(1s): 919-931; DOI: 10.25258/ijddt.16. 919-931