Molecularly imprinted polymers are materials that can capture target analytes using molecular recognition. These polymers can efficiently detect trace target chemical species in biological samples, such as urine and blood, containing various matrix components. We aimed to analyze the interaction of molecularly imprinted polymers with the target molecule lidocaine, which is an amino amide pharmaceutical. Lidocaine‐templated molecularly imprinted polymers were synthesized by bulk polymerization. Gas chromatography‐mass spectrometry showed there was a strong interaction between lidocaine and the recognition site in the molecularly imprinted polymers. Non‐selective adsorption of lidocaine on the molecularly imprinted polymers was confirmed and characterized by Scatchard analysis. The washing solution was optimized for the separation of lidocaine from the structurally similar anesthetics procaine and tetracaine by taking advantage of differences in binding strength. The molecularly imprinted polymers could adsorb lidocaine in urine samples and were not affected by interfering compounds. Consequently, they could be applied to the analysis of biological materials. These results suggest that the molecularly imprinted polymers can be used for separation and detection with high selectivity and could be applied in forensic and clinical analysis.