The main objective of this study was to manufacture an oral rinse using the natural antibacterial agent (phytosphingosine, Doosan, Korea) for the prevention of periodontal disease and dental caries. Phytosphingosine is known to inhibit the growth of bacterial strains and induce apoptotic cell death in human cancer lines. In this study, antibacterial activity and cytotoxicity of oral rinses were performed with an experimental group containing phytosphingosine(PS) in vitro. Control groups consist of two Korean products and two American products containing chlorhexidine and cetylpyridinium chloride, respectively. There was no significant difference between experimental and control groups in the antibacterial activity and cytotoxicity except for Chika Chika Liq (p<0.05). According to the results, antibacterial activity of oral rinse containing PS was 99.62%, the strongest contact inhibition of Streptococcus mutans strain among tested groups. In the cytotoxicity test of oral rinses, PS had a weaker cytotoxicity than control groups in mouse and human normal cell lines and showed the strongest cytotoxicity in human oral cancer cell lines (KB cell). From the results, PS may be widely used as an oral rinse for the healthy and the patients with oral cancers in the near future.