Objective To develop and validate a rapid, robust, and reliable reversed-phase high-performance liquid chromatography (RP-HPLC) method using Analytical Quality by Design (AQbD) principles for the simultaneous estimation of ferrous ascorbate and folic acid in bulk and pharmaceutical dosage forms. Significance This method addresses the need for a straightforward, accurate, and reproducible analytical technique for quality control of ferrous ascorbate and folic acid, ensuring precise measurement in combined formulations and supporting regulatory compliance for pharmaceutical analysis. Methods The RP-HPLC method was systematically optimized for chromatographic parameters, including detection wavelength, flow rate, and mobile phase composition, to achieve excellent separation and peak symmetry of both analytes. Method validation followed ICH Q2(R1) guidelines, evaluating system suitability, accuracy, linearity, and robustness. System suitability parameters such as retention time, peak area, theoretical plates, and tailing factor were assessed. Accuracy was determined at 50%, 100%, and 150% of target concentrations, and linearity was established over wide concentration ranges for both analytes. Robustness was tested by varying flow rates. Results • Consistent system suitability parameters were observed for both analytes, with retention times and peak characteristics within acceptable limits. • The method demonstrated high accuracy, with %RSD values well below 2% and mean recoveries between 98–102% at all tested concentrations. • Linearity was confirmed with correlation coefficients (r²) greater than 0.999 for both ferrous ascorbate and folic acid across their respective ranges. • The method proved robust, as deliberate flow rate changes did not significantly affect retention times or resolution, and all parameters remained within specified limits.Conclusions the AQbD-based RP-HPLC technique is validated as a simple, accurate, linear, and robust method suitable for routine quality control and quantitative analysis of ferrous ascorbate and folic acid in bulk and pharmaceutical formulations.