Diabetes mellitus, associated with α-glucosidase, has been considered as a chronic metabolic disorder, seriously affecting human health. Thus, searching natural α-glucosidase inhibitors and investigating their inhibition mechanism are urgently important. In this study, sixty-two essential oils (EOs), derived from aromatic plants, were found to exert different inhibition on α-glucosidase. The further study revealed that the most potent EOs against α-glucosidase were chuan-xiong, fructus cnidii, sacha inchi, aloe, ganoderma lucidum spore and ginger with IC50 values of 3.02, 2.88, 7.37, 5.06, 5.32 and 7.40 μg/mL. Moreover, the inhibitory mechanism and kinetics studies found that chuan-xiong and sacha inchi were reversible and mixed-type inhibitors. Fructus cnidii, aloe, ganoderma lucidum spore and ginger were reversible and uncompetitive-type inhibitors. It is suggested that EOs, being of natural origin, would be promising anti-α-glucosidase agents.