Objective: Brahmi Ghrita (BG) is a medicated ghee formulation, comprises of Brahmi (), Ela Bacopa monnieri () and Puran ghrita (10 years old cow ghee). It is recommended to treat broadly described mental Elettaria cardamomum illnesses especially in Unmad (Psychosis) and Apasmar (Epilepsy). However, Puran ghrita is not available in ample quantity and in current practice medicated ghee formulations are prepared with fresh go-ghrita (cow ghee). Though the potential of BG is having specific target action on brain tissue, thus to produce desired effect it should be prepared with certain standard operating procedures. Thus before initiating study, validation of manufacture process and in vivo laboratory analysis of Brahmi Ghrita prepared from Puran ghrita and fresh go-ghrita is essential. Materials and methods: In accordance to ayurvedic standard protocols, two samples of Brahmi ghrita were prepared with Puran Ghrita (BG I) and fresh cow ghee (BG II). Further validation and comparative physico-chemical analysis was done for both samples. The physico-chemical parameters such as specific gravity, moisture content, acid value, iodine Results: value, saponification value and peroxide value of BG I and BG II samples were almost alike. The process Conclusion: of manufacture and analytical values of BG I and BG II samples are validated and established in the study.