The fruit cyanidin (Cyn) and pelargonidin (Plr) contents in 73 Fragaria accessions were assayed using a modified HPLC method, and % Cyn (Cyn as % of Cyn-plus-Plr) values were calculated. Plr content increased during fruit ripening, while Cyn content held steady. Consistent with previous studies, % Cyn was comparatively low (5-20%) among twelve F. ×ananassa cultivars, but was >40% in diploid F. vesca. Importantly, three F. chiloensis accessions had greater than 50% Cyn, and ssp. lucida CFRA1691 had the overall highest Cyn content: 707 μg/g FDW. An F1 population of ‘Bountiful’ × ‘Pink Panda’ segregated for both Cyn and Plr contents. Our investigation identifies useful germplasm, offering promise for increasing Cyn content and antioxidant potential through introgressive breeding.