The medicinal plants are believed to be a potential source of medicinal phytochemicals and have a great potential for discovery of new drug candidates. In the Ayurvedic medical system, Barleria prionitis Linn, also known as Kate Koranti, is a well-known medicinal plant. Barleria prionitis Leaves was exhaustively defatted using petroleum ether (BP-PE) and extracted successively with chloroform (BP-CH) and methanol (BPME) using Soxhlet apparatus. Methanolic extract was found potent hence fractionation of methanolic extract was done by column chromatography. Fraction of methanolic extract of Berleria prionitis were subjected to phytochemical screening for tannins, glycosides, steroids, terpenoids, flavonoids, and alkaloids. Isolation of phytoconstituent compound by using High Performance Thin Layer Chromatography (HPTLC). characterization of phytoconstituent was done by using FT-IR, IR, NMR, LC-MS and mass spectrum.