Three stereoselective HPLC methods have been developed for the chiral separation of the enantiomers of three intermediates in the preparation of (+)-tanikolide. The enantiomers were separated on Chiralpak IC (250 × 4.6 mm, 5 μm) in normal phase HPLC. Three intermediates were all baseline separated (RS = 2.84, 2.58 and 3.58, respectively). By comparing the chromatograms of racemates and single enantiomers of the three intermediates, the e.e. values of the single enantiomers were determined and calculated.