Humans have a basic need for better health, yet the accelerating spread of dangerous microorganisms and their detrimental impact on human health present a tremendous challenge to modern science. In this study, piperazine-based cationic oligomeric surfactants (dimeric and tetrameric quaternary ammonium salts) were synthesized. Elucidation of chemical structures of the synthesized cationic surfactants took place using Fourier Transform Infrared, Proton and Carbon Nuclear Magnetic Resonance Spectra. Also, their surface parameters as critical micelle concentration, effectiveness, maximum surface excess and minimum surface area were determined. The tested compounds were evaluated against Gram (+) bacteria as Staphylococcus aureus and Bacillus cereus; Gram (-) bacteria as Escherichia coli and Pseudomonas aeuroginosa. The antimicrobial efficiency was measured via inhibition of the bacterial growth represented as the diameter of the inhibitory zone (mm/mg sample) in contrast to Kanamycin for (G+) bacteria and Ampicillin for (G-) bacteria as standards. The results showed that the prepared cationic oligomeric surfactants can adsorb and orient themselves efficiently at the air/water interface and they have considerable antimicrobial activity against tested species.