*3.3. Purification of the Extract*

The aqueous extract was applied to an Amberlite® XAD-4 column (250 g XAD-4 resin 20–60 mesh; Sigma-Aldrich, MO, USA; 4 × 30 cm column) [3,12], washed extensively with water (1 L) and the saponins eluted sequentially with MeOH (450 mL), acetone (350 mL) and water (250 mL). The MeOH, acetone and water eluates were concentrated, dried, and redissolved in 5 mL of MilliQ water. Finally, the aqueous extract was partitioned with 5 mL isobutanol (v/v). The isobutanolic saponin-enriched fraction was either stored for subsequent mass spectrometry analyses or concentrated to dryness and the components of the extract were further purified by HPCPC. The profile of fractions was also monitored by TLC.
