The extraction of polyphenols from red rice bran was investigated and the purification conditions of red rice polyphenols were established by selecting macroporous resin and eluent concentration. Additionally, the major components of red rice polyphenols were identified, and the effect of red rice polyphenol on the related digestive enzyme activities and glucose adsorption during in vitro small intestinal digestionas well as adsorption of carbohydrate was evaluated. The results showed that the adsorption and desorption rates of red rice polyphenol were relatively high when using macroporous resin HPD400A, and the desorption rate of red rice polyphenol could reach 0.97 with ethanol at a concentration of 70%. The main contents of red rice polyphenol are proanthocyanidins. Red rice polyphenol could inhibit the activities of α-amylases, α-glucosidase, maltase, and sucrase, with 50% inhibitory concentration (IC50) values of 3.61 μg/mL, 2.81 mg/mL, 5.48 mg/mL, and 6.55 mg/mL, respectively. Furthermore, the higher the concentration of red rice polyphenols was, the stronger the inhibitory effect became. Additionally, red rice polyphenols could inhibit in vitro intestinal glucose uptake; the higher the concentration of red rice polyphenols was, the stronger the inhibitory effect became. At the concentration of 2.01 mg/mL, the inhibition rate of red rice polyphenols against glucose uptake was 72.32%.