Prosopis cineraria are small tree generally found in dry arid regions of Arabia and India mainly in Rajasthan. Its state tree of Rajasthan. The flavonoids contained in Prosopis cineraria barks were, identified and characterized. The extraction of bark of prosopis cineraria by using sequential soxhelation, various solvents is used according to polarity Petroleum ether, Chloroform, Ethyl-aceatate, Methanol and water. The metabolic extract was subjected to phytochemical screening. TLC Thin layer chromatography and HPLC for isolation of compound and characterization of isolated compound was done by UV, HPLC and IR. onthe basis of comparisons of spectral data of isolated compound with respects to spectral data of standard rutin. The result show maximum yield of the flavonoids (17.6 gm) was obtained from methanolic extract; phytochemical screenings were done by various specific tests of flavonoids. The Rf value (0.62) of isolated compound has been compared with Rf value of standard rutin. Rf value of isolated compound 4.41 in mobile phase ACN: Water with 0.5 formic acid at lamda max 272. Characterization of isolated compound was done by UV, HPLC,IR ,on the basis spectral analysisstructure was elucidated as 2-(3,4 di hydroxyl phenyl)-5,7 di hydroxyl-3-[α-L-rhmnosyl-(1-6)-βDglucopyranoyloxy]-4H hydroxyl chromen-4-one, rutin was isolated for the first time from this plant (Prosopis cineraria).