The current study focuses on analytical approaches, which involve isolating active components from the Rheum palmatum plant family Polygonaceae. This study emphasises that Rheum palmatum plant was subjected to fractionalization by the column chromatography. During the optimization of the chromatography conditions, various assays were conducted with a gradient of Hexane: Ethyl acetate as the mobile phase.The eluted fractions were run in TLC mobile phase with the solvent Hexane: ethyl acetate ‘7:3’. The fraction showed Rf value equal to standard Aloin in TLC were combined and crystallized. The characterisation technique like UV-visible Spectroscopy and FTIR (Fourier Transform Infrared spectroscopy)are used to portrait that, the probable compound may be the substitute of Aloin.