Abstract: Thin-layer chromatography (TLC) using molecularly imprinted polymers (MIPs) as chiral stationary phases (CSPs ) was applied to the determination of enantiomers of mandelic acid and its derivatives. In preparation of MIPs, L-mandelic acid, L-2-chloromandelic acid and L-4-chloromandelic acid were employed as templates; acrylamide (AM) and ethylene glycol dimethacrylate (EGDMA) were used as functional monomer and cross-linker respectively. With the development system of acetonitrile-5%acetic acid, the racemates of templates were completely separated on the CSPs, the chiral separation factor α were 1.45, 1.62 and 1.56, respectively. Furthermore, the CSPs were able to resolve the racemates of template’s derivatives. The relationship between the chemical structure of the analytes and chiral recognition of the CSPs was also investigated. This method provides a rapid, sensitive and convenient way of analyzing and determining the enantiomers of chiral compounds.