Characterization of a rare triterpenoid and minor phenolic compounds in the root bark of Anisophyllea dichostyla R. Br.

Anisophyllea dichostyla R. Br. (Anisophylleaceae), is a small shrub which grows widely in regions of the Democratic Republic of Congo (DRC) where its root barks are used in folk medicine for the treatment of many debilitating diseases. In a previous work [Khallouki, F., Haubner, R., Hull, W.E., Erben, G., Spiegelhalder, B., Bartsch, H., Owen, R.W., 2007. Isolation, purification and identification of ellagic acid derivatives, catechins and procyanidins from the root barks of Anisophyllea dichostyla R. Br. Food and Chemical Toxicology 45, 472–485] on this species, an appreciable number (16) of phenolic antioxidants (3.32 g/kg) such as ellagitannins (27%) and polyhydroxyflavan-3-ols (catechins and procyanidins; 73%) were isolated and identified. Two fractions, as well as containing minor phenolic compounds also showed evidence of a secondary plant substance similar to a triterpenoid. Following purification of the triterpenoid by semi-preparative HPLC, and recrystallization, the structure was elucidated as bryonolic acid as evinced by comprehensive spectroscopic analyses including 1H and 13C NMR, DEPT, COSY, ROESY, HMQC, HMBC, HPLC–ESI-MS and GC–MS experiments. Bryonolic acid, which is extremely rare in nature, is therefore reported in the family Anisophylleaceae for the first time. Furthermore, the following minor phenolic compounds namely tyrosol, 2-(3-methoxy, 4-hydroxyphenyl)-ethanol, vanillin, syringaldehyde, vanillic acid, syringic acid, gallic acid and ferulic acid were also identified by GC–MS in this species for the first time.