Hemidesmus indicus Linn. is used in the Ayurveda system of medicine in India. Co-exposure to D-galactosamine and lipopolysaccharide is employed in experiments to mimics human viral hepatitis in rats but also damages other organs as D-galactosamine depletes uridine triphosphate pool and inhibits protein synthesis, whereas lipopolysaccharide induces inflammation. We investigated physicochemical properties of extract of Hemidesmus indicus root and its protective potential against multiple organ injury in rats. Doses of Hemidesmus indicus (50, 100 and 200 mg/ kg) were orally administered to different groups of rats before co-exposure to D-galactosamine (300 mg/kg) and lipopolysaccharide (50 µg/kg) intraperitoneally. Hematology, serology, and tissue biochemistry were performed. Aspartate aminotransferase, alanine aminotransferase, bilirubin, cholesterol, urea, uric acid, and creatinine were increased in serum; aniline hydroxylase, lipid peroxidation and cholesterol level in liver, kidney, brain, testis, and spleen tissues were increased. Serum albumin, glucose, and red blood cells; superoxide dismutase, catalase, and glycogen in tissues were decreased after co-exposure to D-galactosamine and lipopolysaccharide. Physicochemical characterization indicated particle size of extract about 43 nm, presence of caffeic acid, quercetin, naringenin and trans-cinnamic acid, and presence of carboxylic acid, aromatic compounds, alkenyl and alkyl groups, alkenes and atmospheric CO2 in Hemidesmus indicus root extract. It was concluded that prophylactic treatment of root extract of Hemidesmus indicus root extract protects multiple organs from injury in dose dependent manner by maintaining diagnostic indices towards control due to its excellent antioxidant potential.