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In the present study, the chemical standardization of unripe fruit of bael {Aegle mormelos (L) Corr.} was assessed preliminary phytochemical screening of bael fruits show the positive test for alkaloids, flavonrides, glycosides, tannins, saponins, steroids and triterpenoids. Major constituents of fruit is mucilage and minerals a coumarin. The preliminary HPTLC studies revealed that the solvent system ethyacetate : methanol: formic acid (7:2:1) was ideal for the 50% ethanolic extract and gave well resolved peaks of crude extract of bael. The numbers of peaks and the Rf values along with the area under the curve are summarized along with the HPTLC profile of 50% ethanolic extract of bael. The 50% ethanolic extract of bael is used for preparation, identification and isolation of active fraction from beutenol fraction of unripe fruit. On the basis of % yields and isolation of review on phytoconstituents, n-butenol fraction was loaded. Preparative TLC was performed in silica gel 60 F254 preparative TLC plates. Individual band were scrapped from the preperative TLC plates and extracted with methanol and dried under reduced pressure. Compound was subjected for spectral analysis and characterization .1HNMR spectroscopy, Mass spectroscopy and IR spectroscopy of spectral analysis shows that active constituent is β-sitosterol.