Background: An Ayurvedic herbomineral formulation containing Shilajit, Swarna bhasma, and other ingredients intended to be used as a rejuvenator for increasing strength stamina and endurance was investigated for its subacute oral toxicity in Wistar rats. Materials and methods: The study was conducted as per the Organization for Economic Cooperation and Development (OECD) guidelines. Test item (TI) was administered to three groups at three dose levels (250, 500, and 1000 mg/kg body weight BW) for 28 consecutive days. Animals in one group were taken as high-dose satellite reversal along with one control group and one satellite control group that received Milli-Q water (10 mL/kg BW). Any clinical sign of toxicity/mortality was observed. Signs of delayed onset of toxicity was observed in satellite groups for the next 14 days. Results: No mortality or clinical sign of toxicity was observed. All animals from the treated and satellite groups showed similar weight gain and feeding as of control group during dosing and recovery periods. Hematological and biochemical parameters were within the range of normalcy in treated and control groups. Necropsy examination did not reveal any significant gross pathology changes. Organ weights in treated group were comparable to control group. Histopathology of high-dose-treated animals and control animals was also comparable. Conclusion: Tested formulation containing Shilajit, Swarna bhasma, and other ingredients was found to be nontoxic at tested doses in both the sexes of Wistar rats on subacute administration. Results of the current study show that TI has no observed adverse effect level (NOAEL) of 1000 mg/kg BW on subacute oral exposure to Wistar rats for 28 days.