Background: Jawarish-e-Amla Sada (JAS) is a traditional Unani formulation commonly used for the clinical treatment of Zof-e-Meda (weakness of the stomach), Zof-e-Kabid (weakness of the liver), Zof-e-Qalb (weakness of the heart), Khafqan (palpitation), Nafkh-e-Shikam (flatulence in the stomach) and Is-Hal-e-Safrawi (biliary diarrhoea). It is a semisolid polyherbal preparation of six medicinal plants. Aims and Objectives: In the present research work an attempt has been made to establish the physicochemical, phytochemical and safety profile of JAS which may be useful for its quality control and standardization. Materials and Methods: Standardized operating procedure for the preparation of JAS was developed in accordance with National Formulary of Unani Medicine. The formulation was also subjected to preliminary phytochemical and physicochemical evaluation. Chemical characterization of the formulation was carried out on the basis of gallic acid content using a validated high-performance thin layer chromatographic method. Safety of the formulation was affirmed by conducting acute oral toxicity in mice. Results: Preliminary phytochemical, physicochemical and chromatographic profile of the formulation was established. The formulation was found safe up to an oral dose of 2 g/kg body weight in mice. Conclusion: Findings of the present work can be used as a reference standard by quality control/assurance laboratory of a pharmaceutical firm in order to have a proper quality check over its preparation and processing.