Investigating 11 Withanosides and Withanolides by UHPLC–PDA and Mass Fragmentation Studies from Ashwagandha (Withania somnifera)

Withania somnifera (WS), also known as ashwagandha or Indian ginseng, is known for its pharmacological significance in neurodegenerative diseases, stress, cancer, immunomodulatory, and antiviral activity. In this study, the WS extract (WSE) from the root was subjected to ultrahigh-performance liquid chromatography with photodiode array detection (UHPLC–PDA) analysis to separate 11 withanoside and withanolide compounds. The quantification validation was carried out as per ICHQ2R1 guidelines in a single methodology. The calibration curves were linear (r 2 > 0.99) for all 11 compounds within the tested concentration ranges. The limits of detection and quantification were in the range of 0.213–0.362 and 0.646–1.098 μg/mL, respectively. The results were precise (relative standard deviation,