LC-HRESIMS Metabolomic profiling of Dalbergia sissoo (Roxb.) ex DC leaves ethanol extract resulted in the tentative characterization of 111 active constituents belong to different chemical classes. Among which four different classes of flavonoid were detected for the first time in the plant in addition to previously reported neoflavonoid and isoflvonoids, with qualitative and quantitative variation. They comprised twenty-one amino acids & other nitrogenous chemicals, five coumarins, eight phenolic acids, five anthocyanins, two alkaloids, and 46 flavonoids. Additionally, the stilbene glycoside E-3,4,5'-Trihydroxy-3'-glucopyranosylstilbene was found in the D. sissoo extract. Along with primary metabolites (Cytidine-3', 5'-cyclicmonophosphate), there are three growth regulators ((+/-)-cis, trans-abscisic acid, beta-indoleacetic acid, and Gibberelin A3). D. sissoo ethanolic leaf extracts successfully protected rabbits from infectious diarrhea when given orally. This is demonstrated by weight parameters, blood measures (Hb, Lymphocytes, Monocytes, and Eosinophil) and antibacterial effect against both gram +ve and gram -ve bacteria. The ethanolic leaves extract of D. sissoo significantly prolonged the onset of diarrhea and reduced the number of wet and total stools at doses of 300 mg/kg and 500 mg/kg as compared to the negative control so, it is highly recommended to use D. sissoo as a natural antidiarrheal drug.