Separation and purification of bioactive botrallin and TMC-264 by a combination of HSCCC and semi-preparative HPLC from endophytic fungus Hyalodendriella sp. Ponipodef12

Two dibenzo-α-pyrones, botrallin ( 1 ) and TMC-264 ( 2 ) were preparatively separated from crude ethyl acetate extract of the endophytic fungus Hyalodendriella sp. Ponipodef12, which was isolated from the hybrid ‘Neva’ of Populus deltoides Marsh × P. nigra L. using a combination of high-speed counter-current chromatography (HSCCC) and semi-preparative HPLC. Botrallin ( 1 ) with 74.73 % of purity and TMC-264 ( 2 ) with 82.29 % of purity were obtained through HSCCC by employing a solvent system containing n -hexane–ethyl acetate–methanol–water at a volume ratio of 1.2:1.0:0.9:1.0. It was the first time for TMC-264 ( 2 ) to be isolated from this fungus. TMC-264 ( 2 ) showed strong antimicrobial and antinematodal activity, and botrallin ( 1 ) exhibited moderate inhibitory activity on acetylcholinesterase.