Abstract High-speed countercurrent chromatography (HSCCC) was applied, for the first time, to the separation of Annonaceous acetogenins. The two major acetogenins of the seeds of Annona atemoya, squamocin and rolliniastatin-2, were isolated in one step using a biphasic mixture of heptane/ethyl acetate/methanol/water. In addition, this method yielded four pairs of highly purified acetogenins: squamocin/molvizarin, rolliniastatin-2/asimicin, neoannonin/atemoyin and isodesacetyl uvaricin/desacetyluvaricin. These pairs were further resolved by I-IPLC, leading to a total of eight pure acetogenins in two steps. Isolated acetogenins, which are homologous, position and configurational isomers, have been identified by comparison with standards and by spectroscopic techniques.
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