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In the present study, we characterised 25 compounds in the extract of Nitraria Tangutorum Bobr. fruits by using the HPLC/MS/MS method. We determined the antioxidant activities of ethanol, petroleum ether, chloroform (CE), ethyl acetate, n-butanol, and water extracts of N. Tangutorum fruits. All extracts were found to exhibit the antioxidant activity and significantly enhance the activities of antioxidant enzymes in H2O2-induced HepG-2 cells. Furthermore, the anti-inflammatory effect and molecular mechanism of the CE of N. Tangutorum Bobr. fruits were evaluated using the dextran sulphate sodium (DSS)-induced ulcerative colitis (UC) mouse model. CE significantly restored the body weight reduction, colon shortening, DAI elevation, and histological score in DSS-induced UC mice (P < 0.01). In addition, CE could remarkably decrease the levels of MPO, IL-1β, TNF-α, and IL-6 and increase the levels of superoxide dismutase, catalase, glutathione, and IL-10 in the colon of DSS mice (P < 0.01). According to western blot analysis, CE inhibited the MAPK/NF-κB signalling pathway activation. The results indicated that CE exerts a protective effect on DSS-induced UC mice, which may be related to the MAPK/NF-κB signalling pathway activation. In conclusion, CE can be used as a therapeutic drug for UC.