Eight new steroids, designated mantuoluosides A-H (1–8), were obtained from the enrichment of steroids of the Datura stramonium L. using HPD-BJQH macroporous resin. Their respective structures were elucidated based on spectroscopic methods and comparison data with literature. The anti-inflammatory activities of these compounds were evaluated on (LPS)-induced RAW264.7 cells with inhibition ratio. It was found that most isolates showed potential anti-inflammatory activity with varying degrees of inhibition rate in a concentration-dependent manner at non-cytotoxic concentrations. [Display omitted] • Eight new steroids, mantuoluosides A-H, were isolated from Datura stramonium L. • The new structures were elucidated by extensive analysis of spectroscopic data. • All steroids exhibited NO inhibitory activities in a concentration-dependent manner. [ABSTRACT FROM AUTHOR]