Introduction: Mrityunjaya Rasa is a herbo–mineral formulation mentioned under Jwara Chikitsa in Rasendra Sara Sangraha along with different Anupanas. It is having ingredients like Shudha Hingula, Shudha Gandhaka, Shudha Vatsanabha, Shudha Tankana, Pippali and Maricha which show significant result on various types of fever. Aim: To prepare Mrityunjaya Rasa as per guidelines given in classical text book Rasendra Sara Sangraha and to validate physico-chemical analysis of the sample. Methods: The preparation of Mrityunjaya Rasa was carried out at practical hall, Department of Rasashastra and Bhaishajya Kalpana, Shri Dharmasthala Manjunatheshwara College of Ayurveda, Kuthpady, Udupi, Karnataka. It was subjected to analysis on parameters like organoleptic characters, loss on drying, total ash, acid insoluble ash, water soluble ash, pH, alcohol soluble extractive value, and water soluble extractive value. Results: The results of Analytical study showed specific characteristics which are in accordance with the properties of Mrityunjaya Rasa. Conclusion: The study showed significant results, which proves quality of the drugs and efficacy.