A procedure is proposed for separation and preconcentration of lanthanium, cerium and yttrium using octacarboxymethyl-C-methylcalix resorcinarene impregnated onto a polymeric matrix, Amberlite XAD-16. The optimization process was carried out using 23 factorial design and three variables (pH, concentration of metal ion, sorption flow rate) were regarded as factors. For Ce3+ extraction, the variance analysis revealed that pH and concentration of Ce3+ was statistically significant. However, for La3+ and Y3+ pH was significant only. The precision of the method (%RSD) was 2.27 for La3+, 1.97 for Ce3+ and 2.01 for Y3+ at 1 µg mL−1 for ten replicate analyses. The preconcentration factors of La3+, Ce3+ and Y3+ were 125, 83 and 100 respectively. The robustness of this procedure is demonstrated by the recovery achieved for determination of La3+, Ce3+ and Y3+ in the presence of several cations. The procedure was satisfactorily applied to the separation of La3+, Ce3+ and Y3+ from each other and also from common actinides such as UO22+ and Th4+ by sequential acidic elution system.