Ascophyllum nodosum is a brown seaweed that grows abundantly in the US Northeast coastal region. This study examined the seasonal variation of A. nodosum in phenolic contents and subsequent antioxidant, α-glucosidase and α-amylase inhibitory activities. A. nodosum was harvested monthly and extracted in hot water and the resulting extracts were spray-dried. The results indicate a clear seasonal variation in terms of phenolic content, with June and July being the highest (36.4 and 37 mg/g, respectively) and May the lowest (21.8 mg/g). The antioxidant activities, in terms of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, correlated with the phenolic contents observed (r = 0.81), with the month of July being the highest (58%) and April the lowest (26%). Similarly in terms of Trolox equivalent, July had the highest activity (15.53 μM) and April and May the lowest (8.40 and 8.27 μM, respectively). α-glucosidase inhibitory activity exhibited a pattern similar to the phenolic contents observed with July having the highest inhibitory activity (IC(70) 2.23 μg) and April the lowest (IC(70) 26.13 μg), resulting in an inverse correlation between IC(70) values and total phenolic content (r = -0.89). Such seasonal variation is believed to be caused by temperature-related stress considering that A. nodosum is a cold water species.