Mate tea (MT) is a rich source of phenolic compounds that vary depending on geographical origin and mode of preparation. Total phenolic concentration and antioxidant capacity of Mate tea ( Ilex paraguariensis) products were determined in this work. In addition, a representative MT was tested for in vitro inhibition of human colon carcinoma cell proliferation. Total polyphenol concentration, was measured using Folin–Ciocalteau method, ranged from 90 to 176 mg gallic acid eq (GAE)/g dry leaves (DL) in traditional MT and from 40 to 113 mg GAE/g DL in MT added with other flavouring ingredients. It was estimated that a cup of tea (250 ml) containing one teaspoon (5 g) of instant MT could provide an average intake of 1.5 g GAE. Fresh tea (FT) from Mate leaves displayed high antioxidant capacity (85 ± 1%) and preferentially inhibited 50% of net growth of human colorectal adenocarcinoma cells CaCo-2 (GI 50 = 1.0 ± 0.03 μg/ml) and HT-29 (GI 50 = 105.2 ± 15.2 μg/ml) when compared with the CCD-33Co normal colon fibroblast cell line (GI 50 > 300 μg/ml). MT inhibited in vitro colon cancer cell proliferation possibly mediated via pro-oxidant activities, therefore represents a potential source of chemopreventive agents that deserve further investigation.