Study On The Non-iridoid Chemical Constituents Of Gentiana Macrophylla Pall.

Gentiana macrophylla Pall.(G.macrophylla)is a perennial herb,belong to Gentianaceae family and Gentiana genus Sect.Cruciata Gaudin.As one of the four sources of traditional Chinese herbal medicine "Radix Gentianae Macrophyllae",the dried roots of G.macrophylla are used for the treatment of rheumatism and rheumatoid arthritis.The common bioactive components of Gmacrophylla are iridoids,and there is lack of systematical and complete studies on other potentially active components,such as flavonoids,triterpenoids,steroids and alkaloids.The study aims to isolate non-iridoids from chloroform extraction phase of 70%ethanolic extract of G.macrophylla's dried roots.Column chromatography and high performance liquid chromatography,coupled with thin layer chromatography,were used for repeated separation and purification.1D-NMR(1H,13C and DEPT135)and 2D-NMR(HSQC,HMBC and H-H COSY),HR-ESI-MS,CD and other Spectroscopic methods were used to analyse the structure of the pure compounds.A total of 16 compounds were identified,4-Hydroxyl-roburic acid methyl ester(1),Swerpalactone(2),Swermacrolactone(3),6R,8R)-hydroxyl-Swerimus-lactone A(4),(6R,8S)-hydroxyl-swerimuslactone A(5),Blumenol C(6),9-Epi-blumenol C(7),6-Demethoxycapillarisin(8),N-tetraco-sanoylanthranilic acid(9),N-pentacosanoylanthranilic acid(10),N-hexacosanoylanthranilic acid(11),N-heptacosanoylanthranilic acid(12),N-octacosanoylanthranilic acid(13),Swerimusic acid B methyl ester(14),3-Hydroxyl-gentioxepine(15),5-Hydroxymethylisochromen-1-one(16).All the 16 compounds are non-iridoids,and compounds 1,4,5,14 and 15 are news,2,3,8,16 are firstly isolated from Gentiana.Lipopolysaccharide(LPS)was used to stimulate the production of an inflammatory factor NO,in mouse RAW264.7 cells to establish an inflammation model in vitro,to study whether the five new compounds have anti-inflammatory activity.The inflammation model was dosed with the five new compounds of different concentrations to observe the inhibitory effect of production of NO,aspirin as a positive control.The results revealed that all the five new compounds showed some anti-inflammatory activity.And compound 15 showed stronger activity than the positive control,which provided theoretical basis and data support for further studies on its activity.