Withania somnifera, from the Solanaceae family also known as Ashwagandha, Indian ginseng or Winter cherry is an important medicinal plant being used in Ayurvedic and indigenous medicines. Its pharmaceutical role has been of great interest to the scientific community. The plant produces secondary metabolites, withanolides, produced via oxidation of steroids and therefore has a steroid backbone bound to lactone and lactone derivatives. Among withanolides, Withaferin-A, has shown potential as an anti-inflammatory, anti-cancerous and antioxidant agent. Withania somnifera is well distributed all over India and is found growning under different climatic conditions. The present study showcases phytochemical screening of Withania somnifera plants on the basis of Withaferin-A concentration. Germplasms collected from different regions of India were used to determine the environmental variation on phytochemical constituents. The study involved the standardization of extraction procedures to derive the maximum extraction efficiency and used methanol extraction, homogenization and sonication platform. Extracts of leaves and roots were analyzed using HPLC and the concentration of withaferin-A in leaves was found to be much higher than roots and there were variation in its concentration between different germplasms. This study showed that withaferin-A can be extracted to high yield and that yield varies with different geographical locations.