Pedneault, K., Dorais, M., Léonhart, S., Angers, P. and Gosselin, A. 2014. Time-course accumulation of flavonoids in hydroponically grown Achillea millefolium L. Can. J. Plant. Sci. 94: 383-395. In recent decades, the use of plant-based medicines as health products has increased considerably all over the world. As greenhouse hydroponic culture allows standardized cultural methods to be used, it may be valuable for reducing the risks associated with harvesting medicinal plants from the wild, such as species dissemination, species misidentification, adulteration, and non-hygienic handling, while allowing the production of high yields of clean, standardized biomass year-round. To evaluate the potential of hydroponic culture for medicinal plant production, the present study investigated the accumulation patterns of apigenin, luteolin, apigenin glycosides, and the chlorogenic acid 5-caffeoylquinic acid in the plant organs of A. millefolium at five phenological stages from 35 to 102 d after sowing, and drew a comparison with outdoor-grown plants at 122 d after sowing. The results showed two flavonoid accumulation peaks: one at the early growth stage (35 d after sowing) and one at early flowering (87 d after sowing). At 87 d after sowing, most of the apigenin glycosides were concentrated in the roots (3.80% wt/wt, dry weight basis), whereas free apigenin and luteolin were located mainly in the flower heads (1.25 and 0.86% wt/wt, dry weight basis, respectively). Early flowering was the best harvesting stage for optimal flavonoid production in terms of active compounds per plant and kilograms of plant biomass per cultivated area. At 122 d after sowing (phenological stage 4), the outdoor-grown plants were nine times smaller than the early flowering plants (87 d after sowing) from the hydroponic system and had a root-tissue apigenin glycoside level that was five times lower than that of the hydroponically grown plants. In conclusion, the use of a hydroponic growing system reduced by 29% the time required to reach phenological stage 4, which corresponds to maximum plant bioactive concentration, in comparison with field production. Therefore, hydroponic culture represents an effective alternative to outdoor production and can result in standardized, high-quality medicinal plant biomass with potential flavonoid yields approximating 515 mg per plant.