The air-dried and milled flowers, fruits,leaves, and twigs of I. spicata (1.2 kg) were extracted by maceration with MeOH (4 × 3 L) at room temperature overnight.
After removing the solvent under reduced pressure, the combined and concentrated methanol extract (129 g) was suspended in a mixture of 90% MeOH−H2O (1 L), then partitioned with hexane and CHCl3, in turn, to afford hexane- (5 g) and CHCl3 (32 g)-soluble extracts.
Bioactivity-guided fractionation was performed on the cytotoxic CHCl3 extract of the flowers, fruits, leaves, and twigs of I. spicata (IC50 values against HT-29, 697, and Raji cells of 4.3, 2.2, and 5.0 μg/mL, respectively), leading to the isolation of 10 compounds