Analytical Evaluation and in Vitro Cytotoxic Activity of Arbudahara Rasa

Background: Arbudahara Rasa is a classical herbo-mineral formulation described in Ayurvedic literature for the management of Arbuda (tumor-like conditions). Scientific validation of such formulations through analytical and biological studies is essential for establishing quality, safety, and therapeutic potential. Objective: To evaluate the physicochemical characteristics, instrumental analytical profile, and in vitro cytotoxic activity of Arbudahara Rasa. Materials and Methods: Arbudahara Rasa was prepared according to classical Ayurvedic procedures involving Shodhana, Bhavana, and Puta processes. The finished formulation was subjected to organoleptic evaluation, Bhasma Pareeksha, physicochemical analysis including Loss on Drying (LOD), Total Ash, Acid Insoluble Ash, Water Soluble Ash, pH determination, and mercury estimation. Instrumental characterization was performed using Fourier Transform Infrared Spectroscopy (FTIR) and Inductively Coupled Plasma–Optical Emission Spectroscopy (ICP–OES). Cytotoxic activity was assessed against HepG2 human hepatocellular carcinoma cell lines using the MTT assay and compared with Doxorubicin as a standard drug. Results: The formulation appeared as a light brown, fine powder without characteristic odour or taste. Classical Bhasma Pareeksha tests such as Rekhapurnatva, Varitaratva, and Unnama were positive. Physicochemical analysis revealed LOD 1.717% w/w, Total Ash 92.00% w/w, Acid Insoluble Ash 55.46% w/w, Water Soluble Ash 10.87% w/w, mercury content 491 ppm, and pH 9.05. FTIR analysis demonstrated the presence of both organic phytoconstituents and inorganic mineral phases, while ICP–OES confirmed mercury incorporation within the formulation. MTT assay showed concentration-dependent cytotoxic activity against HepG2 cells with an IC₅₀ value of 202µg/mL. Maximum inhibition of 65.76% was observed at 320µg/mL. Conclusion: Analytical findings confirmed the physicochemical stability and successful incorporation of mineral and herbal constituents in Arbudahara Rasa. The formulation demonstrated moderate dose-dependent cytotoxic activity against HepG2 cell lines, suggesting potential anticancer properties that warrant further pharmacological and clinical investigation.