Approximately 7.4 kg of dried bark was pulverized
and extracted with EtOH then further partitioned4
to
yield 545.5 g of F005 [brine shrimp lethality test (BST)
LC50 ) 1.5 íg/mL]. From this, 500.5 g was loaded onto
a Si gel column and eluted with hexane and increasing
percentages of CHCl3, then CHCl3 and increasing
percentages of MeOH. In all, 60 fractions were collected, and fractions 30-36 were combined (21.67 g).
This material was separated on successive open columns
packed with Si gel. Repeated HPLC of fractions active
in the BST5
yielded compounds 1-4.