Introduction: Rasaushadhies (metal and mineral) preparations are unique preparations that include Bhasma, herbo-mineral preparations and Kupipakwa rasayana hold a significant place in Ayurvedic pharmaceutics. Nearly 70 % of formulations include a combination of one or more metallic/mineral with several herbs which have a supporting role in improving efficacy, relieving symptoms of the disease and to achieve long and healthy life. Shilasindur is a one of the Kupipakwa rasayana containing mercury, sulphur and arsenic disulphide and indicated in skin disorders, respiratory tract disorders and other infectious diseases. Aim and Objectives: Evaluation of repeated dose (Sub chronic) toxicity study of Shilasindur along with its characterization and antimicrobial activity. Materials & Methods: Shilasindur will be prepared in three batches as per classical reference and by using Electric Muffle furnace. The prepared formulation will be assessed with Organoleptic characters, physicochemical parameters and ICP – AES (Inductively coupled plasma atomic emission spectroscopy), FTIR (Fourier-transform infrared spectroscopy), XRD (X-Ray Diffraction). Sub chronic toxicity study will be carried out according to the OECD guidelines 408 and 452 as well as CCRAS guidelines of safety and toxicity.  Epsilometer test (E-test) will be used to check antimicrobial activity of Shilasindur. Results: The analytical parameters will be assessed to establish the pharmaceutical standardization. Toxicity study will be done in three doses to ensure its safety. Statistical analysis will be done by applying unpaired t-test and ANOVA test. Conclusion: If significant positive results obtained in this work then it will be a valuable contribution and evidence that the drug Shilasindur is safe for consumption at treatment doses.