Abstract: Background and Objective: Elaeagnus latifolia Linn., commonly known as wild olive, has been used as a traditional medicine to treat a variety of ailments, including anticancer, antioxidant, free radical scavenging activities and as an astringent. The study aimed to isolate and characterize the bioactive components from the flowers of Elaeagnus latifolia. Materials and Methods: The dried coarsely powdered flowers were extracted with water for 48 hrs and subjected to chromatography, characterised by UV-Visible, FT-IR, MS, 1H NMR and 13C NMR. Results: Phytochemical screening revealed the presence of glycoside, phytosterols and saponins. 1H NMR has revealed a signal at 7.44 ppm for O-H3 proton, the signal at 8.7 ppm for H26. Six methyl protons appeared at 0.81, 0.95, 0.97, 1.04, 1.29 and 1.32 ppm for H29, H21, H27, H26, H18 and H19. 13C NMR has given signal at 134.7 and 121.4 ppm for C5 = C6 double bond, 71.7 ppm for C3 β-hydroxyl group, 19.4 ppm for angular methyl carbon atom for C19, 29.6 ppm for C25, 36.6 ppm for C20, 56.5 ppm for C17, 28.8 ppm for C16, 42.3 ppm for C13, 40.2 ppm for C12, 50.1 ppm for C9, 32.0 ppm for C2, 37.2 ppm for C1. The purification of the compound resulted in white crystals with melting points 140-146°C and molecular mass 414.71. Conclusion: Based on the spectral data analysis and chemical reaction, the isolated compound was concluded as β-sitosterol. This compound has not been previously isolated from the flowers of Elaeagnus latifolia.