Musa balbisiana fruits is an important and commonly used plant for its medicinal and pharmacological properties. Urolithiasis has become a very common clinical disease in over the world. The present study aimed to evaluate in vitro anti-urolithic, antiinflammatory, and antibacterial activities of crude and fractionated extracts of M. balbisiana fruits. The M. balbisiana fruit extracts were evaluated anti-urolithic property by nucleation and aggregation assays of calcium oxalate crystallization, anti-inflammatory activity by protein denaturation assay and red blood cell membrane stabilization, and antibacterial activity using the well diffusion agar method, minimum inhibitory concentration assay, and minimum bactericidal concentration assay. Results indicated that crude extract and different fractions had anti-urolithic, antibacterial, and anti-inflammatory activities. Chloroform fraction had the highest ability against calcium oxalate crystals formation with an IC50 value of 22.04 ± 1.09 µg/mL, which was comparable with sodium citrate (202.08 ± 18.46 µg/mL). In the active in against crystals aggregation, n-hexane fraction had the highest activity with an IC50 value of 414.77 ± 10.85 µg/mL, whereas IC50 value of sodium citrate was 1390.33 ± 92.66 µg/mL. Furthermore, crude and fractionated extracts presented anti-inflammatory activity through inhibition protein denaturation and red blood cells membrane stabilization. Simultaneously, the crude extract and different fractions exhibited the ability to against E. coli, P. aeruginosa, and S. aureus. Taken together, the present study had demonstrated that M. balbisiana fruits has properties that support urolithiasis disease. [ABSTRACT FROM AUTHOR]