Abstract
An efficient, accurate, sustainable, and reproducible chromatographic method for Quercetin estimation was developed, optimized, and further validated using a Design of Experiments approach in compliance with ICH guidelines. The Box-Behnken Quality-by-Design optimized critical parameters, yielding robust performance, excellent linearity (R(2) = 0.9996, 0.05-50 μg/mL), and high sensitivity (LOD: 40.27 ng/mL, LOQ: 122.03 ng/mL). Deliberate modifications evaluated by analysis of variance verified the method's significance (P < 0.0001). The method utilises a Phenomenex HyperClone C18 LC Column (250 × 4.6 mm; 5 μm, 130 Å) with an isocratic mobile phase consisting of acidified water (0.1% v/v Triethylamine, pH 3.0 ± 0.05) and Acetonitrile (55:45% v/v). Forced degradation studies revealed that quercetin was stable under mild acidic, photolytic, and oxidative conditions at room temperature but suffered significant degradation under basic, strong acidic, and overall accelerated conditions (60 °C for 6 h), indicating sensitivity to harsh environments. Greenness evaluation results show an AGREE score of 0.6 and a favorable ComplexGAPI profile, highlighting minimal energy consumption during the 12-min run and moderate solvent/reagent use. The method's efficiency and repeatability promote robust quercetin quantification, with wide applications in various pharmaceuticals and nanomaterials, as demonstrated by its effective use in silver-based Metal-Organic Frameworks in this study.