Background: Many of the vivid colours found in fruits, vegetables, flowers, and even tea are caused by interesting natural substances called flavonoids. Because they appear to provide mild, protective effects that assist our systems in daily ways, these plant-made compounds have become a hot issue in research and health. Flavonoids shield plants from oxidative stress, illnesses, UV rays, and herbivores. E. neriifolia, or Euphorbia neriifolia. There are several ethnomedical applications for E. neriifolia. In addition to treating whooping cough, gonorrhea, leprosy, asthma, dyspepsia, jaundice, enlarged spleen, tumors, bladder stones, gastrointestinal issues, and leucoderma, the latex of E. neriifolia is used as a laxative, purgative, rubefacient, carminative, and expectorant. Brittle, hot, carminative, and useful for treating bronchial infections, tumors, aches, inflammations, and stomach swellings, leaves also improve appetite. The goal of the current study was to identify and isolate the unique flavonoid found in Euphorbia neriifolia leaves. Methodology: Extracts showing presence of flavonoids will be subjected to column chromatography for further separation of individual compounds. Further characterization of isolated compound was carried out by chromatographic and spectral analysis. Result: Based on current finding the compound C1 was identified as quercetin, a flavanol compound containing phenolic hydroxyl group and conjugated aromatic rings. Conclusion: The present study successfully isolated and characterized flavonoid compounds from Euphorbia neriifolia using chromatographic and spectral techniques.