A new molecularly imprinted polymer (MIP) has been synthesized for the selective extraction of trans,trans-muconic acid (ttMA) from urine samples, followed by high-performance liquid chromatography analysis with ultraviolet detection. The synthesis was based on non-covalent interactions, and 4-vinylpyridine was used as a functional monomer. The analytical calibration curve was prepared using a pool of five urine samples of non-smokers spiked with ttMA standards with concentrations that ranged from 0.3 to 10 mg L⁻¹ (r² = 0.999). The limit of quantification was 0.3 mg L⁻¹ (lower than the biological exposure limits suggested by the ACGIH). The within-day and between-day precision and accuracy presented relative standard deviations and relative errors of less than 15%. The analytical frequency was 4 h⁻¹ (considering extraction and separation/quantification steps), and the same MIP cartridge was efficiently used for approximately 100 cycles. All figures of merit were similar or better than those obtained by the procedure based on ionic exchange extraction. The proposed method could be an interesting alternative for the routine analysis of ttMA in urine for biological monitoring procedures of human exposure to benzene.