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A new megastigmane derivative, (6R,9S)-6′-(4″-hydroxybenzoil)-roseoside 1 and two known compounds, the biflavone agathisflavone 2 and the carboxylic acid derivative 4-hydroxibenzoic acid 3 were isolated and purified from leaves and stems of Ouratea polyantha Engl. Agathisflavone was isolated in a single high-speed counter-current chromatography run, meanwhile the megastigmane and the carboxylic acid derivatives were purified in two steps, a combination of high-speed countercurrent chromatography and analytical column chromatography. All structures were elucidated on the basis of spectral evidences and comparisons with literatures. Compound 1 was characterized by [α]D20, UV-Vis, IR, MS, 1H-NMR, 13C-NMR, HMQC, HMBC, COSY and NOESY. Compounds 1 and 2 were tested as hepatic glucose-6-phosphatase inhibitors.

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