The N-Dopa tablet is a proprietary Ayurvedic poly-herbal formulation widely used in Parkinson's disease, chronic fatigue syndrome, Myalgia and Muscular dystrophy. This study aims to standardize the formulation using advanced analytical techniques and pharmacopoeia standards. The physicochemical testing, heavy metal analysis and microbiological limit test evaluations were carried out in accordance with Ayurvedic pharmacopoeia of India. All the heavy metals were determined to be within the acceptable ranges. The extractive value revealed increases in water soluble extractive, which suggests higher bioavailability in a water medium. In addition, the microbial load was found to be safe for ingestion by humans in terms of microbes. This assessment could aid in determining the drug's authenticity and provide pertinent data for the safer and more effective use of this formulation in therapeutics. These methods will assist drug manufacturers in adhering to regulations and substantiating the stability, safety, and therapeutic efficacy of their goods.