HPLC-based phytochemical characterization of ashwagandha Pratap [Withania somnifera (L.) Dunal] cultivated in the soils of Haridwar, Uttarakhand

Abstract

This study aimed to evaluate the phytochemical composition of Withania somnifera (L.) Dunal ‘Pratap’ variety cultivated in the soils of Haridwar, Uttarakhand, with a focus on quantifying key withanolides using High-Performance Liquid Chromatography (HPLC). Soil samples from the cultivation site at Teliwala nursery were collected and analyzed using standard APHA protocols and Atomic Absorption Spectroscopy to assess physicochemical and micronutrient profiles. Plant parts such as roots, stems, and leaves were subjected to HPLC-DAD analysis on
a Shimadzu LC-2030C 3D system for quantification of six major withanolides, including withaferine A, withanoside IV & V, withanolide A & B, and withanone.
The soil was moderately alkaline (pH 8.37) with high organic carbon (1.43%), and low nitrogen (2.79 mg/kg), and phosphorus (7.0 mg/kg), though sulphur and zinc were deficient. The leaf tissues exhibited the highest total withanolide content (10.285 µg/mg), particularly withaferine A and withanone, followed by stems (1.358 µg/mg) and roots (0.886 µg/mg). The distinct withanolide profiles across plant parts highlight tissue specific accumulation trends. Notably, withanoside IV dominated in stems, while withanoside V peaked in leaves. The study demonstrates a clear association between favorable soil conditions and enhanced secondary metabolite synthesis in W. somnifera. While roots remain commercially important, the phytochemical richness of leaves suggests underutilized pharmacological potential. Future work should explore soil metabolite correlations across varied agro-ecological zones to optimize medicinal quality and cultivation strategies.