This study aimed at examining the antibacterial activity of R. nasutus ointment in different concentration of S. aureus bacteria growth by in vitro dilution method. The antibacterial activity of R. nasutus ointment was determined based on minimum inhibitory concentration (MIC) value and minimum bactericidal concentration (MBC) from R. nasutus ointment against S. aureus bacteria. The results showed R. nasutus ointment that the MIC value was 250,000 μg/mL up, whereas the MBC was 125,000 µg/mL. The variation of R. nasutus ointment concentration significantly has a relation with S. aureus bacterial growth. These results indicated that R. nasutus ointment potentially used as an alternative medicine for skin disease because it has an ability as antibacterial.