DEVELOPMENT AND OPTIMIZATION OF A QBD-GUIDED

RPHPLC METHOD FOR THE QUANTITATIVE DETERMINATION OF RUTIN

IN BULK AND LIPID–POLYMER HYBRID NANOCARRIERS

Quality by Design (QbD) is a structured, scientifically driven methodology aimed at ensuring product quality through well-defined objectives and proactive risk management. In the present work, a reverse-phase high-performance liquid chromatography (RPHPLC) method was developed and optimized following QbD principles for the precise quantification of rutin in its bulk form, as well as in lipid–polymer hybrid nanoparticle (LPHN) formulations. Optimization was guided by Box Behnken design using Design Expert® software, which enabled systematic evaluation of three critical analytical parameters. The developed method demonstrated linearity within a concentration range of 10–50 μg/mL. System suitability testing showed a theoretical plate count of 7033 and a tailing factor of 1.12, both within acceptable analytical limits. High precision was confirmed by relative standard deviation (%RSD) values below 1%, while robustness was maintained with variability restricted to under 2%. The quantitative analysis yielded a rutin recovery rate of 98.78 ± 0.61%, with no interference from nearby eluting peaks. Validation of the method adhered strictly to ITH Q2(R1) guidelines, confirming the method’s suitability across key parameters, such as accuracy, linearity, precision, robustness, and specificity. This QbD-driven development approach provided a clear insight into variable interactions and enabled the creation of a dependable, reproducible, and accurate analytical method well-suited for application in pharmaceutical formulation development and quality control processes.