HPLC Analysis Of Ashwagandha Supplements Using Alliance™ iS HPLC System

By Fadi L. Alkhateeb, Jinchuan Yang, Paul D. Rainville

This study presents a highly efficient HPLC method for quantifying key withanolides in ashwagandha root and its commercial supplements. Rigorous system suitability testing validated the method's accuracy and precision.

Ultrasound-assisted extraction using a 60:30:10 ethanol:methanol:water solvent for 30 minutes proved optimal for maximizing withanolide recovery from ashwagandha root. The developed method effectively separated and quantified major withanolides in the USP reference root sample.

However, analysis of commercial ashwagandha supplements revealed a variable presence and concentration of withanolides. Many supplements contained only a subset of the withanolides found in the USP root, and their levels were often significantly lower.