This study aimed to assess the volatile components and bioactivity potential of Ruprechita polystachya Griseb cultivated in Egypt. Air dried leaves was extracted was dichloromethane:methanol 1:1. Gas chromatography mass spectrometry (GC-MS) was applied to study the chemical constituents of n-hexane fraction while standard in vitro bioassays were used to detect the cytotoxic, antioxidant and antimicrobial potential of R. Polystachy. GC-MS analysis of n-hexane fraction showed the tentative presence of twenty-eight secondary metabolites classified as follow: ten fatty acid esters, eight hydrocarbones, two diterpenes, one triterpene, one phenyl propanoid, one phenolic derivative, one alkyl phenyl ketone, one α–tocopheroid, one γ-lactone derivative, one ketonic compound and one steroidal compound. The major identified compounds were: Ethenone-1-(2,3-dihydroxy-4-methoxy-6-methyl phenyl) (xanthoxylin) (49.41%), eugenol (4.89%), 9-octadecenoic acid (z)-methyl ester (3.18), hexadecenoic acid methyl ester (2.64%) and 9,12- octadecadienoic acid(z,z)-methyl ester (2.44%). R. polystachya extract showed cytotoxic activity against HeLa, MCF-7, HepG-2, PC3 and HCT-116 cancer cell lines with IC50 10.20, 12.79, 16.10, 26.38 and 28.23 µg/ml resprctively. In antioxidant assay using ascorbic acid as areference drug, R. polystachya extract exhibited a high potentiality with IC50 34.59 µg/ml while the corresponding fractions showed less potentiality. The antimicrobial activity of the extract/fractions was assessed using agar diffusion assay and crude extract showed a good inhibition against C. Albicans with activity index 74.1 % while it showed less activity against bacterial strains, E. Coli and B. Subtilis, with activity indicies 34.6 and 60.9 % respectively. R. polystachya can be considered as a potential source of bioactive compounds and more comprehensive exploration of its bioactive secondary metabolites is recommended.