The aim of the current work was to isolate and characterize structurally fascinating and biologically intriguing compounds from the dried roots of Premna latifolia. Dried plant roots were subjected to soxhalation with ethyl alcohol and later to column chromatography. The individual compounds were isolated by preparative thin-layer chromatography followed by structural characterization using various spectral ways like LCMS, IR, 1D-NMR and 2D-NMR (1H-1H NMR and 1H-13C NMR). Three totally different compounds were isolated and characterised as Compound 1: n-Tridecanyl n-Tetracosanoate, Compound 2: Stigmanstan -3β-olyln-octadec-9ʹ, 12ʹ-dienoate and Compound 3: n-Tetracosanol for the first time from Premna latifolia roots.