Annona glabra L. is known for its edible fruit and it has numerous medicinal and therapeutic benefits. The aim of this study is to evaluate the antimicrobial activity of different solvent extracts of Annona glabra fruits followed by chemical profiling of the most bioactive extracts. The antimicrobial activity was estimated using disc agar plate diffusion test and chemical characterization was achieved using GC-MS analysis. Results revealed that all tested extracts exhibited noticeable antimicrobial activities with remarkable consideration to ethyl acetate, n-butanol and methanolic extracts. Ethyl acetate extract showed antimicrobial inhibition zone values ranged from 10 to 16mm. Whereas, n-butanol extract exhibited antimicrobial inhibition zone values ranged from 10 to 18mm. In addition, the methanolic extract displayed antimicrobial inhibition zone values ranged from 8 to 21mm, all against S. aureus, E. coli, C. albicans and A. niger. GC-MS analysis showed that 3,5-decadiyne, 2,2-dimethyl- (24.93%), hexadecanoic acid, methyl ester (12.92%), and hexadecanoic acid, methyl ester (23.69%), were detected as main chemical ingredients in the ethyl acetate, n-butanol, and methanol extracts, respectively. Annona glabra fruits could be a good source of natural antimicrobial agents.