Inhibitory effects of the Noble muscadine grape extracts and the representative phytochemicals for anthocyanins (i.e., cyanidin and cyanidin-3,5-diglucoside) on two enzymes, that is, α-glucosidase and pancreatic lipase, were investigated regarding their antidiabetic activities. The study demonstrated that the anthocyanin extracts and the selected chemicals obeyed the competitive mode against the enzymes. The methanolic extracts of whole fruit and skin of the muscadine showed inhibitory activities against the α-glucosidase with their IC(50) values at 1.50 and 2.73 mg/mL, and those against the lipase at 16.90 and 11.15 mg/mL, respectively, which indicated that the muscadine extracts possessed strong antidiabetic activities. Particularly, the ethyl acetate (EtoAc) extract and the butanol (BuOH) extract exhibited much higher inhibitory activities against both enzymes than the CHCl(3) and water extracts, while the majority of anthocyanins existed in the BuOH fractions. Moreover, cyanidin exhibited a much stronger antidiabetic activity than cyanidin-3,5-diglucoside, suggesting that anthocyanins may have higher inhibitory activities after being digested. Further chromatographic analysis by high-performance liquid chromatography-mass spectrometry identified five individual anthocyanins, including cyanidin, delphinidin, petunidin, peonidin, and malvidin glycosides.