The objective of the present investigation is to evaluate the antimicrobial potency of the terpenoidfractions isolated from Luffa cylindrica seeds against various pathogenic microbes. The seeds werepowdered and extracted with methanol in soxhlet appratus based on phytochemical screening.Three terpenoid components were isolated by column chromatography and identified by thinlayer chromatography and chemical analysis which were designated as LCSF4*, LCSF6* & LCSF8*respectively. Disc diffusion method was employed to determine the antimicrobial effectiveness oftest compounds I, II and III (LCSF4*, LCSF6* & LCSF8*) against 6 microbial species viz., Staphylococcus (S.)aureus, Bacillus (B.) subtilis, Escherichia (E.) coli, Pseudomonas (P.) aeruginosa, Candida (C.) albicans andAspergillus niger. The disc was saturated with 100 µl of each compound, allowed to dry and introducedon the upper layer of seeded agar plate. The plates were incubated overnight at 37°C. Microbialgrowth was determined by measuring the zonal inhibition diameters. Compound I showedmaximum potency against gram positive S. aureus (21 mm) in comparison with standard ciprofloxacin(38 mm), whereas the same compound was completely devoid of activity against both the fungitested. Compound II was found to be highly sensitive against both the gram negative E. coli (20 mm)and P. aeruginosa (22 mm). Compound II was found to exhibit maximum potency against the fungi C.albicans (15 mm) and A. niger (20 mm). Compound III was found to be very effective against both thegram positive S. aureus (20 mm) and B. subtilis (15 mm) respectively.