A previously unreported secoiridoid glycoside, cruciatoside ( 1 ) was isolated from the aerial parts of Gentiana cruciata L. along with ten known compounds eustomoside ( 2 ), eustomorusside ( 3 ), gentiopicroside ( 4 ), 6'- O -β-D-glucopyranosyl gentiopicroside ( 5 ), loganic acid ( 6 ), isoorientin ( 7 ), isovitexin ( 8 ), isovitexin 2'' -(E)- ferulate ( 9 ), mangiferin ( 10 ), and 2-methyl-inositol ( 11 ). The chemical structures of the isolates were elucidated based on extensive 1 D and 2 D NMR experiments as well as HRMS analysis. All isolates were evaluated for their in vitro anti-inflammatory and analgesic activities. Compounds 9 , 4 , and 7 (200 µM) showed moderate anti-inflammatory activity by inhibiting nitrite production from LPS-induced RAW 264.7 macrophage cells, with the inhibition rates of 39.5%, 25.8% and 22.9% respectively without exhibiting substantial cytotoxicity. Besides, 1 , 2 , 4 , and 7 exerted the highest decrease in IL-6 levels. Moreover, compound 4 showed in vitro analgesic activity by decreasing the PGE 2 level comparable to the reference drugs.