Polyethyleneimine (PEI) and ethylenediamine (EDA) as modifiers were bonded on active carbon (AC) surface for specific selective extraction of quercetin from Oldenlandia diffusa. The characteristics of the modified AC materials that were obtained were investigated by field emission-scanning electron microscopy (FE-SEM) and Fourier transform infrared spectrometer (FT-IR). The interactions between quercetin and the AC materials were investigated by fitting the static adsorption data to four linear and nonlinear adsorption isotherm models. Of these four models, the Langmuir-Freundlich adsorption isotherm was proved the best for investigating quercetin on AC materials. Scatchard analysis was used to evaluate the binding properties of the AC materials for quercetin. Solvent extraction and solid-phase extraction (SPE) were optimized, and the effect of the mobile phase pH was investigated to improve the performance for the separation of quercetin on high performance liquid chromatography (HPLC). The results from the validation of the proposed analytical method demonstrated that the EDA-modified AC was the most suitable SPE cartridge for the purification of quercetin from O. diffusa.