Objective: The present work is aimed to formulate Ashwagandha kalpa and evaluate the impact of fermenting agent.Methods: A. kalpa is a marketed patent proprietary Ayurvedic product prepared by adopting the Ayurvedic principles of madya sandhana and it belongs to the class of formulations known as Sandhana kalpana. It is given to the patients suffering from hypertension, insomnia, paralysis, loss of concentration, etc. In the preparation of madya sandhana, the sandhana dravya (fermenting agents) play a key role. The present study was taken up to see the effect of three different sandhana dravya (fermenting agents), i.e., Dhataki pushpa (DAK), Madhuka (MAK), and yeast (YAK) on the formulation characterization of A. kalpa.Results: The prepared samples were tested for their physicochemical parameters, i.e., total solid, pH, specific gravity, sugar content, alcohol content, refractive index, phytochemical screening, quantitative assay for secondary metabolites, and high-performance thin-layer chromatography (HPTLC) fingerprinting. The findings of phytochemical evaluation, quantitative assay, and HPTLC fingerprint show marked variation among three different prepared samples of A. kalpa. The preliminary phytochemical estimation for the detection of secondary metabolites was done. The study revealed the presence of highest percentage of alkaloids in YAK, tannins in DAK and saponins in YAK. Total solid content was found to the highest in MAK. Physicochemical parameters such as pH, total solid content, refractive index, alcohol content, and specific gravity also showed marked variation. Alcohol content for the DAK and YAK was found same.Conclusion: sandhana dravya (fermenting agent) causes variations in different physicochemical and phytochemical parameters in the formulation and development of A. kalpa.