The aim of the present study was to evaluate the qualitative analysis of phytochemicals and antimicrobial activity of various solvent extracts of Boerhaavia diffusa L. (Family: Nyctaginaceae) leaves. The antimicrobial activity of different solvent extracts of B. diffusa L. leaves were tested against the Gram-positive and Gram-negative bacterial strains by observing the zone of inhibition. The Gram-positive bacteria used in the test were Staphylococcus aureus, Bacillus subtilis, Streptococcus faecalis and Micrococcus luteus, and the Gram-negative bacteria were Escherichia coli, Pseudomonas aeruginosa, Salmonella typhii, Klebsiella pneumoniae, Proteus vulgaris, Serratia marcescens, Shigella flexneri and Vibrio cholerae. It was observed that ethanol, methanol, chloroform, ethylacetate and aqueous extracts showed activity against Gram-positive and Gram-negative bacteria. The ethanol extract of B. diffusa L. leaves showed more activity against Gram-positive (e.g. S. aureus, zone of diameter 11 mm) and Gram-negative bacteria (e.g. E. coli, zone of diameter 9 mm) when compared to other solvent extracts except V. cholerae. The results confirmed the presence of antibacterial activity of B. diffusa L. leaves extract against various human pathogenic bacteria.