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The Croton Macrostachys is used for the treatment of malaria, venereal diseases, cough,diabetes, constipation, tape worms and hepatitis. Following this, several compounds have been isolated and identified from the different parts of the plant. The present study is designed for the extraction, isolation and structural elucidation of compounds from methanol extract of the root bark of Croton Macrostachys. TLC has been used for the identification of the various components. Column chromatographic separation of the crude methanol extract using different solvent system with increasing polarities resulted in isolation of various compounds. Of those isolated compounds, with appropriate amount, structural elucidation was performed for two of them (compound CM-6 and CM-10) using spectroscopic techniques: UV-Vis, IR, 1H NMR, 13C NMR, DEPT-135 and 2D NMR (COSY, HSQC, and HMBC).