Hexane, ethyl acetate and methanol extracts of whole plant of Boerhavia diffusa were screened for phytochemical and biological activities. Qualitative phytochemical screening via colorimetric method and the quantitative estimation of phenolic and flavonoid content were performed. Antioxidant assay using DPPH scavenging method was studied. Antimicrobial screening of plant extracts was done by cup diffusion technique. Cytotoxic activity of B. diffusa was studied by brine shrimp bioassay and anthelminthic activity was evaluated in vitro in Pheretima posthuma. This study revealed B. diffusa as a source of various phyto-constituents such as alkaloids, glycosides, saponins, tannins, carbohydrates, cardiac glycosides, flavonoids and terpenoids. Quantitative estimation of total phenol was found to be maximum in BEE i.e. 29.73±0.88, BME 19.8±2.02 and in BHE 9.15±0.304 mgGAE/g. Similarly, the total flavonoid content was found to be 17.44±0.75 in BEE, 14.43±0.23 in BHE and 3.678 mg QE/g in BME. Ethyl acetate extract showed its antibacterial activity against all tested pathogens except Escherichia coli whereas Staphylococcus aureus and Salmonella Typhi were resistant to methanol and hexane extract. The zone of inhibition (ZOI) of ethyl acetate extract against S. Typhi and B. cereus was found to be 18 mm and 14 mm respectively. The MIC value of BEE in S. Typhi was 3.125 µg/ml and in B. cereus was 12.5 µg/ml. The preliminary screening of anticancer property of B. diffusa i.e. BSLT in methanol was found to be 165.19 µg/ml. B. diffusa was also found to contain anthelmintic property. The study helped in further exploration of medicinal properties of B. diffusa by phytochemical screening and biological activities paving the path for study and investigation in this plant.