Fig 1.
Sarsasaponin is composed of a hydrophilic sugar moiety (a glycoside, coloured blue) and a lipophilic steroidal moiety (a sapogenin, coloured black) joined through a glycosidic bond (purple) to make an amphipathic molecule that produces a persistent foam when shaken in water.
Fig 2.
Fisher projections of smilagenin and sarsasapogenin.
The smilagenin molecule, on the left, displays the numbering system of each carbon atom. The glycoside was attached at the C3 position but after hydrolysis is occupied by a hydroxyl group (here, attached by a purple bond). At the other end of the rings is C25, a chiral atom bound to a methyl group (C27) in the R-configuration for smilagenin or the S-configuration for sarsasapogenin (in red).
Table 1.
Some physical properties of smilagenin and sarsasapogenin.
Fig 3.
The composition of Smilax spp, root [13].
Fig 4.
The photograph on the left shows dilutions of tincture after they were shaken vigorously for 15 seconds. The photograph on the right was take 10 minutes after the tubes were shaken. Tube B is the same tube in both photographs, but it has lost about half a millilitre of foam during the 10-minute interval, leaving 3.5mls of persistent foam.
Fig 5.
A volume of marc (plant material) was mixed in 3 volumes of menstruum. In metric units that is 480 grams of powder (specific gravity of 0.48) in 3 litres of 45% ethanol (specific gravity of 0.93). The concentration of marc retained in solution must be determined by measuring the dry weight of the tincture.
Table 2.
Batch quality of four batches of tinctures.
Table 3.
Results at four time points (top row) from four reaction conditions.
Fig 6.
Yields from tincture incubated at 80 oC in 6N HCl for 1, 2, 4 and 8 hours (left) and in 2N HCl for 2, 4, 8 and 16 hours (right).
Colours correspond to sarsasapogenin (blue), smilagenin (orange) and non-sapogenins (grey). The circled values above each column are the total sapogenins (sum of sarsasapogenin and smilagenin).
Fig 7.
Yields from tincture incubated at 70 oC in 6N HCl (left) and 2N HCl (right) for 2, 4, 8 and 16 hours.
Colour coded as in Fig 6.
Table 4.
Changes in maximum yields at 80 oC or 70 oC and 6N or 2N HCl.
Fig 8.
Photographs of separatory funnel containing chloroform and unhydrolyzed tincture.
A) Chloroform gently added to Mock Reaction Mixture settled below it. B) When mixed, an emulsion formed. C) An hour later, most of the chloroform, now tinted, had returned to the bottom and the emulsion lost its colour. D) Several hours later, after the emulsion had dispersed, the chloroform was collected, leaving this aqueous layer containing saponins.
Fig 9.
Photographs of separatory funnel containing chloroform and hydrolysed tincture.
A) Aqueous (top) and chloroform (bottom) layers, ready to be collected. B) Aqueous layer remaining after chloroform layer has been collected, displaying “dark disk” that forms during extraction of hydrolysed tincture.
Fig 10.
Comparison of methods for saponin preparation.
Top diagram details the protocol used by Jacob and Simpson, with the values they reported. Bottom diagram displays our protocol for comparison.
Fig 11.
Comparison of methods for hydrolysis and collection of sapogenins.
Top diagram details the protocol used by Jacob and Simpson, with the values they reported. Bottom diagram displays our protocol for comparison.
Fig 12.
Demonstration of oil solubility in solutions of ethanol and water.
The photograph on the left was taken by ambient light and the photograph on the right was taken with the assistance of a flash.