Berries are a rich source of various polyphenols. The objective of this study was to investigate the bioavailability of polyphenols from berries. Middle-aged subjects (n = 72) consumed moderate amounts of berry or control products for 8 weeks in a randomized, placebo-controlled dietary intervention trial. Average intake of berries was 160 g/day (bilberries, lingonberries, black currants, and chokeberries). Plasma and urine polyphenols were analyzed by GC-MS and HPLC and berry polyphenols by HPLC. The total intake of polyphenols was 837 mg/day. Plasma quercetin, p-coumaric acid, 3-hydroxyphenylacetic acid, caffeic acid, protocatechuic acid, vanillic acid, homovanillic acid, and 3-(3-hydroxyphenyl)propionic acid increased significantly from the baseline in the berry group compared to the control group (p < 0.05). The urinary excretion of quercetin, p-coumaric acid, and 3-hydroxyphenylacetic acid increased significantly in the berry group compared to the control group (p < 0.05). In conclusion, a number of polyphenols are bioavailable from a diet containing moderate amounts of blue and red berries.