The aim of this work was to investigate the content and the composition of alkylresorcinols (ARs) in different wheat and barley cultivars, and in fractions obtained by progressive pearling. Three commercial winter wheat cultivars, characterized by different hardness and technological quality, and three barley cultivars, including hulled and hull-less types, were selected. Two different protocols of sequential pearling were applied, one for wheat and hull-less barley and another one for hulled barley. Pearling of wheat and hull-less barley cultivars gave five fractions (each corresponding to 5% of original grain weight) and 75% of the residue. In the case of hulled barley eight pearled fractions and 60% of inner kernel were obtained. In wheat ARs were prevalently located in the 5-10% intermediate fraction, while for barley results varied depending on the cultivar. In the hull-less cultivar, the AR content progressively decreased from the outermost fraction (0-5%) towards the inner layers, while for hulled barley the highest AR content was observed in the 10–15% fraction, evidencing lower amounts in the coarse hull (included in the 0-5% and 5-10% fractions). Based on the different localization of ARs in the cereal kernel, progressive pearling can be employed to obtain enriched fractions that could be used to enhance ingredients and products rich in these bioactive compounds.