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The Argentinean Patagonia berries <i>Berberis microphylla</i>, <i>Berberis darwinii</i>, and <i>Fragaria chiloensis</i> ssp. <i>chiloensis</i> f. patagonica were investigated for their polyphenol content and composition by means of liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry. The in vitro antioxidant activity and inhibition of metabolic syndrome-associated enzymes (&#945;-glucosidase, &#945;-amylase, and lipase) of the fruit extracts was assessed. The most complex polyphenol profile was found in the <i>Berberis</i> samples, with 10 anthocyanins, 27 hydroxycinnamic acids, 3 proanthocyanidins, 2 flavan-3-ol, and 22 flavonols. <i>Fragaria</i> presented four anthocyanins, nine ellagitannins, two proanthocyanidin dimers, one flavan-3-ol, and five flavonols. The <i>Berberis</i> samples showed the best antioxidant capacity, while <i>Fragaria</i> displayed better activity against &#945;-glucosidase and lipase. The phenolic content and composition of the Argentinean Patagonia berries was similar to that reported for Chilean samples but with some chemical differences between Eastern (Argentina) and Western (Chile) Patagonia. The data obtained supports the consumption of these berries as sources of beneficial polyphenols.