This study aimed to evaluate the changes in the content of functional components and antioxidant activity of rice and barley according to the milling process used. A considerable amount of γ-oryzanol was observed in unmilled rice and barley. However, γ-oryzanol was not detected in the rice and barley after the milling process. The highest content of γ-aminobutyric acid (GABA) was observed in the unmilled Keunalbori-1-ho barley. The contents of biotin in all samples also decreased in the milled grains compared to the unmilled grains. The highest content of total polyphenols and flavonoids was observed in the Heuksujeongchal barley, and the highest radical scavenging activity was also found in this grain. The milling process led to a decrease in the content of functional components, including γ-oryzanol, GABA, biotin, polyphenols, and flavonoids in both rice and barley. These results may be useful in the development of processed foods using cereal grains.