Phenolic contents of barley and malt extracts and their corresponding antioxidant activities were investigated using a chromatographic online antioxidant detection system. Ethyl acetate extracts of barley and malt were separated using reverse phase HPLC and compounds eluting from the column were submitted to two UV–visible detections: one for the phenolic compounds; and the other for the reduced form of the radical cation 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+) after the compounds were allowed to react online with it. Prodelphinidin B3 and procyanidin B3 were identified as two major contributors in the antioxidant activity of barley, in addition to catechin. Malting had a dramatic impact on these three compounds by resulting in a sharp decrease in their detected amounts and the associated antioxidant activities. Two other antioxidants, ferulic and sinapic acids, showed a better ability to withstand not only malting but also brewing steps. As for the overall phenolic content and antioxidant capacity, the study showed that malting allowed a better release and/or extraction of phenolic compounds, while the first brewing step caused the most significant damage by drastically decreasing the total polyphenols and their activity. Hopping however did not significantly affect neither the phenolic content nor the antioxidant activity.