Amomum Tsaoko Crevost et Lemarie is a kind of spices and traditional Chinese medicines with great commercial value. Traditional treatment often uses essential oils from Amomum Tsaoko in aromatherapy. In this study, researchers analyzed the composition of steam-distilled essential oil from Amomum tsaoko (ATEO) and evaluated its potential antioxidant and anti-inflammatory activity. GC–MS identified a total of 33 volatile compounds, the majority of which were alcohol (1,8-cineole and nerol) and aldehyde ((2E)-2-decenal) compounds. Additionally, ATEO showed great anti-inflammatory activity in LPS-induced RAW 264.7 macrophages. ATEO markedly inhibited the nitric oxide (NO) release, and decreased the expression level of TNF-α, IL-1β, IL-6 and monocyte chemotactic protein-1 (MCP-1) in a dose-dependent manner. Furthermore, ATEO significantly reduced the expression of iNOS, COX-2, NF-κB p-p65 and p-ERK. These results suggested that ATEO has great anti-inflammatory activity in LPS-induced RAW 264.7 cells, which will facilitate the development of ATEO as a functional food.