A study on the separation of lithium isotope was carried out with a 1,16-dioxa-4,7,10,13- tetraazacyclooctadecane-4,7,10,13-tetramerrifield peptide resin [N4O2·4M]. The resin having N4O2 as an anchor group has a capacity of 3.8 meq/g. Upon column chromatography [0.15 cm (I.D)×29 cm (height)] using 0.01 M NH4Cl as an eluent, the single separation factor, α=1.038 was obtained by the Glueckauf theory from the elution curve and isotope ratios.