*3.2. Extraction of Saponins*

The saponins were extracted as described previously [3,12]. Briefly, the visceral masses were removed, freeze dried (VirTis, BenchTop K, New York, NY, USA) and pulverized to a fine powder using liquid nitrogen and a mortar and pestle. The pulverized viscera sample was extracted four times with 70% ethanol (EtOH) (400 mL) and filtered using Whatman filter paper (No. 1, Whatman Ltd., Maidstone, England, UK) at room temperature. The extract was concentrated under reduced pressure at 30 ◦C using a rotary evaporator (Büchi AG, Flawil, Switzerland) to remove the ethanol, and the residual sample was freeze-dried. The dried extract was dissolved in 400 mL of 90% aqueous methanol (MeOH), and partitioned against 400 mL of *n*-hexane twice. The water content of the hydromethanolic phase was then adjusted to 20% (v/v) and then to 40% (v/v) and the solutions partitioned against CH2Cl2 and CHCl3, respectively. The hydromethanolic phase was concentrated and then freeze-dried. The dried powder was solubilized in 10 mL of MilliQ water (18.2 MΩ, Millipore, Bedford, MA, USA) in readiness for chromatographic purification.
