The direct determination of both the enantiomeric purity and absolute configuration of timolol maleate has been accomplished by a rapid, specific, and sensitive 1H-NMR spectroscopic method. The (R)-(+)- and (S)-(-)-enantiomeric resonances were effectively resolved in CDCl3 solution using (R)-(-)- or (S)-(+)-2,2,2,-trifluoro-1-(9-anthryl)-ethanol as a chiral solvating agent. Enantiomeric impurities were determined on the basis of the intensities of the methyl protons resonances; the assignment of enantiomeric configurations was based on the relative field position of these resonances and the prediction from molecular models. Based on the analysis of synthetic enantiomeric mixtures the results correlated well with the values for the enantiomers added and with that previously found by 1H-NMR with a chiral Pr(III) shift chelate. However, the present approach shows less reliance on reagents and solvents of high purity, and does not require strict anaerobic working conditions.