Anthocyanin-rich acetone extracts of barley grain of four hulless pigmented genotypes were investigated to determine their potential as functional food ingredients. The purple barley cultivar contained 11 anthocyanins, whereas only one anthocyanin, peonidin derivative, was observed in blue, black and yellow barley. The total anthocyanin content of pigmented barley genotypes ranged from 3.2 to 678.5 mg kg−1 in whole grain and from 4.5 to 1654.6 mg kg−1 in bran. The purple barley bran extract gave the highest DPPH radical scavenging capacity, superoxide radical scavenging capacity and total antioxidant activity. The half maximal inhibitory concentration (IC50) of angiotensin I-converting enzyme (ACE) of anthocyanin-rich extract of purple barley grain and bran was 8.77 and 4.54 mg mL−1, respectively. Purple barley appears to have great potential uses for the promotion of human health and development of nutraceuticals and functional foods.