Uterotibb is one of the potent Unani medicines that offers a natural solution to various human health issues. The present investigation explores the active phytochemical constituents of Unani medicine Uterotibb using GC-MS analysis. This study further evaluated the antioxidant potential and antimicrobial activity against a panel of human pathogens such as Staphylococcus aureus, Bacillus Subtilis, E. coli, and Klebsiella pneumonia. GC-MS studies revealed the presence of 388 phytochemical compounds in the alkali aqueous extract of Uterotibb. The main active biomolecules were stigmast-5– en-3-0l, and oleate (2.84%). Cyclotrisiloxane, hexamethyl (1.74%). 1,2,5-oxadiazole-3-carboxamide4-amino-N-(2-methoxyethyl)-(1.75%). Arsenous acid (1.48%). Pentane,1,1-thiobis (1.46%). 7,7,9,9,11,11-hexamethyl-3,6,8,10,12,15hexaoxa-7,9,11-trisilaheptadecane (1.46%). 4-(1,1dimethylpropyl-phenol, trimethylsilyl ether (1.43%). 1,2-bis (trimethyl silyl) benzene (1.36%). Adamantane methyl amine (1.32%). 1,4-benzenediole,2,5-bis(1,1 dimethyl ethyl)-(1.29%). 2-chloro aniline-5-sulphonic acid (1.28%); Acetic Acid, nitro-, methyl ester (1.24%) and Caprolactone oxime, (NB)-O-[(diethylboryloxy) (ethyl)boryl] (1.21%). Notably, Uterotibb extract exhibited antimicrobial activity in a concentration-dependent manner, where the highest concentration of 5 mg/mL showed a maximum zone of growth inhibition of 14.95±0.35 mm against Staphylococcus aureus, and the lowest zone of inhibition was 13.26±0.88 mm against Bacillus subtilis at the same concentration. The lowest MIC value was exhibited at 2 mg/mL concentration against Staphylococcus aureus where the highest MIC value of 4 mg/mL was observed against K. Pneumoniae. Further, the antioxidant potential of the extract exhibited scavenging activity of 67.115 ± 0.05 and 73.67 ± 0.09 using DPPH and ABTS assays, respectively. Finally, GC-MS profiling and bioactivity studies of the present work validate Uterotibb's potential as a natural Unani medicine for human care.