Molecularly imprinted polymers(MIPs)for solid-phase extraction and pre-concentration of esculetin have been successfully prepared by the bulk polymerization method using esculetin as a template molecule.Polymers of varying composition were prepared using different monomers(4-vinylpyridine,methacrylic acid,and acrylamide),ethylene glycol dimethacrylate as the cross-linker,2,2-azobis(2-methylpropinitrile)as the initiator,and different porogen solvents(ethanol,acetone/methanol,and acetonitrile).The best polymer was obtained when 4-vinylpyridine was used as the monomer and acetone/methanol(3:2)as the porogen solvent,whereas the template:-monomer:-cross-linker ratio was 1:4:20.The imprinting factor of the selected MIPs for esculetin was 3.77.The polymers were evaluated according to their selective recognition properties for esculetin and structurally-related compounds(esculin,scopoletin,coumarin,and 7-methoxycoumarin).Chemical and morphological characterizations of the polymers were investigated by FTIR and scanning electron microscope,which confirmed a high degree of polymerization.Surface area,pore volume,and pore size of the polymer were investigated by Brunauer-Emmett-Teller analysis.MIPs were also successfully used as solid-phase adsorbent materials for the extraction of esculetin from tobacco leaves.Esculetin contents in dried tobacco leaves were found to be(9.27±0.17)μg g-1. Molecularly imprinted polymers (MIPs) for solid-phase extraction and pre-concentration of esculetin have been successfully prepared by the bulk polymerization method using esculetin as a template molecule. Polymers of varying composition were prepared using different monomers (4-vinylpyridine, methacrylic acid, and acrylamide, ethylene glycol dimethacrylate as the cross-linker, 2,2-azobis (2-methylpropinitrile) as the initiator, and different porogen solvents vinylpyridine was used as the monomer and acetone / methanol (3: 2) as the porogen solvent, the the template: -monomer: -cross-linker ratio was 1: 4: 20. The imprinting factor of the selected MIPs for esculetin was 3.77 The polymers were evaluated according to their selective recognition properties for esculetin and structurally-related compounds (esculin, scopoletin, coumarin, and 7-methoxycoumarin). Chemical and morphological characterizations of the polymers were investi gated by FTIR and scanning electron microscope, which confirmed a high degree of polymerization. pore area, pore volume, and pore size of the polymer were investigated by Brunauer-Emmett-Teller analysis. MIPs also also successfully used as solid-phase adsorbent materials for the extraction of esculetin from tobacco leaves. Esculetin contents in dried tobacco leaves were found to be (9.27 ± 0.17) μg g-1.