Background: It is well-known fact that medicinal plants are the resources of promising drugs for many ailments. Although, a number of research have been conducted on these plants, however, study related to their cytotoxic and antibacterial activity is of paramount important due to worldwide increase microbial resistance to antibiotics. Objective: The objective of present study was to determine the cytotoxicity and antibacterial activities of Cochlospermum tinctorium roots extract against some clinical isolates of bacteria. Water Material and Methods: and methanol were used in extraction of roots by percolation. The antibacterial activity Cochlospermum tinctorium was carried out using agar well diffusion technique against , , , and E. coli Proteus specie K. pneumoniae P. aeruginosa S. aureus, the minimum inhibitory concentration and the minimum bactericidal concentration were also determined using standard procedure The extracts were also subjected to Phytochemical Screening, Brine-shrimp lethality assay, the Lc 50 values were determined by linear regression analysis method. The result of phytochemical Results: screening revealed the presence of secondary metabolites, the result of antibacterial activity ranged from 7.23±0.20mm to 32.67±0.58mm inhibition zones, N-hexane extract have the highest zone of inhibition (32.67±0.58mm) against at 2000μg/ml concentrations, the extract was active on all the tested K. pneumoniae microorganisms at different concentrations, the MIC was lowest for at (62.5µg /ml) and highest for K. pneumoniae S. aureus E. coli K. pneumoniae and at (125µg/ml), the MBC result showed that, the extract was bactericidal to at (62.5 µg/ml followed by at (125µg/ml) and at (250µg/ml), the cytotoxicity of the plant extract (LC50) S. aureus E. coli showed that, the plant was weakly toxic. These findings therefore support the local use of Conclusion: C. tinctorium root extracts for the treatment of bacterial infections in Northern Nigeria.