Novel ciprofloxacin composite imprinted materials are synthesized by using co-precipitation polymerization of dual functional monomers (methacrylic acid and 2-vinylpyridine) and polystyrene-co-divinylbenzene. The intermolecular interactions between monomers and template are evaluated by molecular modeling analysis. The physicochemical properties of the obtained polymers are characterized using FT-IR, TGA, and SEM. Batch adsorption experiments are used to investigate adsorption properties (kinetic, pH, and isotherm). These polymers are employed to prepare the solid phase extraction cartridges, and their extraction performances are analyzed by the HPLC-UV method. DFT calculations indicate that hydrogen bonding and π−π stacking are the driving forces for the formation of selective rebinding sites. The obtained polymers exhibit excellent adsorption properties, including fast kinetics and high adsorption capacity (up to 10.28 mg g⁻¹) with an imprinted factor of 2.55. The Scatchard analysis indicates the presence of specific high-affinity adsorption sites on the imprinted polymer. These absorbents are employed to extract CIP in river water with recoveries in the range of 65.97–119.26% and the relative standard deviation of 3.59–14.01%. Furthermore, the used cartridges could be reused at least eight times without decreasing their initial adsorption capacity.