An extra-facile chiral liquid chromatography-time of flight mass spectrometry (LC-TOFMS) analytical method of amino acid enantiomers has been developed without a derivatization process. The enantioseparation of eighteen proteinogenic amino acids (except for proline) was simultaneously performed using a combination of a chiral column (CROWNPAK CR-I(+)) and a TOFMS within 15 min. An isocratic condition of a simple mobile phase comprising acetonitrile/water/trifluoroacetic acid (96/4/0.5) gave baseline separation of all underivatized amino acid enantiomers on the chiral column. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.