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The floral resins of the genus Clusia L. (Clusiaceae Lindl.) mainly consist of polyisoprenylated benzophenones, a class of substances which is noteworthy due to its broad spectrum of biological activities. This paper describes the isolation of clusianone from the male flowers of Clusia fluminensis Planch. & Triana, a native species from the Brazilian coast, using, for the first time, high-speed counter-current chromatography with a nonaqueous solvent system composed of n-hexane–acetonitrile–methanol (2:1.25:0.5, v/v/v) in 2 hr, with purity higher than 90%.