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The root and flowers of Butea monosperma (Lam.) were extracted with methanol. Extensive chromatographic separation and purification with the organic solvents was done. Four phytochemicals were separated and their structures were established based on various spectroscopic techniques. Isolated crude extract was subjected for antibacterial activity against gram-negative and gram-positive bacteria like Bacillus megatarium, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa using standard protocols with MIC (Bacterial) in range of 7.5 to 25 µg/ml. The antifungal activity was also carried out against strains of Alternaria , Fusarium and Aspergillus flavus within the range between 10 µg/ml to 300 µg/ml MIC (Fungal). The results of antibacterial activity were compared with a standard antibiotic disc of ciprofloxacin and norfloxacin (5 to 30 µg/disc) against the same microbial strains. The results of antifungal activity were compared with nystatin (100 to 200 µg/disc) against the same microbial strains.