Resolution of clenbuterol hydrochloride enantiomers and propranolol hydrochloride enantiomers was achieved by thin-layer chromatography using methyl-β-cyclodextrin impregnated silica gel GF254 plates. The effects of stereoselective auxiliary (acetonitrile/different alcohols) were studied. Resolution of clenbuterol hydrochloride enantiomers could be realized by alcohols such as ethanol, n-butanol, 2-butanol or tert-butanol together with acetonitrile, and propranolol hydrochloride enantiomers could only be resoluted by ethanol or n-butanol together with acetonitrile. The optical conditions of simultaneous resolution were obtained as follows: 15.00g silica gel GF254 plates impregnated with 1.00g methyl-β-cyclodextrin, acetonitrile/n-butanol (V/V = 90:10) mixed solvent as spreading solvent, being resolved under ambient temperature. Under these conditions, RF value of resolution of clenbuterol hydrochloride enantiomers is 3.47, RF value of resolution of propranolol hydrochloride enantiomers is 3.35. The two couples of enantiomers were all achieved base line separation.