The study was carried out to ascertain the functional groups present in Piper longum, and to assess its ability to disrupt biofilms caused by throat infectious methicillin resistant Staphylococcus aureus (MRSA) strains. Ethanol extract of Piper longum was subjected to fourier transform infrared (FT-IR) analysis which revealed the presence of different pharmacologically active functional groups. Two strains of methicillin resistant Staphylococcus aureus (MH4 throat isolate and MTCC 96 standard strains) were used. Minimum inhibitory concentration (MIC) was performed by micro dilution assay on a 96 microtitre well plate (OD 600 nm) and was found to be 1 mg/ml for both isolate MH4 and MTCC 96. This was further confirmed by agar well diffusion assay and growth curve analysis. Biofilm inhibition assay was performed at sub-MIC concentrations using microtitre plate (24 well plates) assay. The highest biofilm disruption was observed at 0.5 mg/ml concentration for both strains. This study revealed potential antibiofilm activity exhibited by the ethanol extract of Piper longum against the ever rising methicillin resistance in Staphylococcus aureus infections.