Figure 3.

Stacked-up HPLC profiles of WSSO and the post-supercritical fluid extraction (SCFE) exhaust of WS seeds, with known standards: Withanolides contents were identified using standards mix (black line). Post-SCFE exhaust of Withania somnifera Dunal (WS) seeds (red line) showed good presence of withanoside IV, withaferine A, withanoside V, and withanolide A, at standard retention times (Rt). WSSO (blue line) chromatogram show identifiable peaks of withferine A (at Rt, 31.86 min) and of withanolide A (at Rt, 38.65 min). Most of the withanolides tested were identified in WS seed exhaust only. However, withanolide B and withanone were not decteced (ND) in any of the test samples. Relevent structures of withanolides are displayed at their identfied peak, as per standard mix. The observed concentrations of withanolides have been listed as % w/w with their respective retention time (Rt) in both test samples.

Sample Name	Withanoside IV Rt 21.96	Withaferin A Rt 31.86	Withanoside V Rt 34.33	Withanolide A Rt 38.65	Withanone Rt 39.32	Withanolide B Rt 46.58	Total Withanolides
WSSO	ND	0.022	ND	0.001	ND	ND	0.023
Post SCFE Exhaust of WS Seeds	0.807	0.359	0.744	ND	ND	ND	1.910