This study aimed to profile the phytochemical components of the hexane and dichloromethane (DCM) extracts obtained from the fresh Egyptian Broccoli leaves (Brassica oleracea L. var. italica), together with their biological activity. The extracts were analyzed by GC-MS, whereby 25 and 12 compounds were obtained from the hexane and DCM extracts, respectively. The major constituents of the hexane extract were 3-methyloctacosane (58.76%) followed by 15-methyltriacontane (15.09%) while the major constituents of the DCM extract were 3,3,17,17-tetraethylnonadecane (33.71%), benzyl isothiocyanate (16.44%) and 13-methylnonacosane (13.38%). The two extracts were investigated for anti-Helicobacter pylori and antimicrobial activities using the agar-well diffusion method. The hexane and DCM extracts possessed significant antimicrobial activities against H. pylori, with an activity index of 1.25 and 1.35, respectively and a minimum inhibitory concentration (MIC) of 7.8 µg/mL for both extracts. The hexane extract showed strong activity on Escherichia coli (ATCC8739) and Staphylococcus aureus (ATCC6538) (with MIC = 62.5 and 31.125 µg/mL, respectively), a moderate activity on Bacillus subtilis (ATCC6633), Klebsiella pneumoniae (ATCC13883) and Candida albicans (ATCC10221) (with an MIC = 15.62 µg/mL) for the three strains. While DCM extract showed a strong activity against S. aureus and E. coli (MIC =1.38 and 1.36 µg/ mL, respectively), a moderate activity on B. subtilis, K. pneumoniae and C. albicans (MIC = 3.9, 62.5 and 7.8 µg/mL, respectively). Both extracts showed a low activity against Aspergillus niger with an activity index of 0.57 and 0.66, respectively.