Lupinifolin from Derris reticulata Craib. was extracted with hexane by Soxhlet extractor and purified by crystallization. The yellow needle-shaped lupinifolin crystals were identified and confirmed by nuclear magnetic resonance (NMR) spectra and Liquid chromatography mass spectrometry (LC/MS). The lupinifolin showed minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 8 and 16 µg/ml against Methicillin resistant S. aureus (MRSA), respectively. The flow cytometry (FCM) was performed to determine the alteration of cytoplasmic membrane (CM) permeability of MRSA by using Propidium iodide (PI) 5 µg/ml as an indicator for bacterial membrane integrity. It was found that the bacterial CM permeability was effected by lupinifolin with the MIC of 8 µg/ml comparable to the control when investigated by Propidium iodide intensity. Additionally, DNA laddering assay was carried out to evaluate apoptosis in bacterial cells. It was shown that the lupinifolin has no effect on DNA fragmentation.