Thanneervittan Nei is a traditional Siddha herbal medicated ghee used for the treatment of gynecological disorders, especially Menorrhagia. The present study was carried out to evaluate the physicochemical properties, phytochemical constituents, and HPTLC profile of the formulation. Physicochemical parameters such as pH, specific gravity, acid value, peroxide value, and saponification value were analysed using standard methods. Preliminary phytochemical screening was performed to identify the presence of active compounds. HPTLC fingerprint analysis was also carried out for standardization of the formulation. The study revealed the presence of important phytoconstituents such as flavonoids, tannins, phenols, alkaloids, and terpenoids. The HPTLC profile showed distinct bands indicating the presence of various bioactive compounds. The results of the study help in the quality evaluation and standardization of Thanneervittan Nei.