Mechanistic study on the high-selectivity enantioseparation of amino acids using a chiral crown ether-bonded stationary phase and acidic, highly organic mobile phase by liquid chromatography/time-of-flight mass spectrometry.

•Chiral crown-ether bonded phase enabled fast separation of chiral amino acids.•All proteinogenic amino acids except proline were resolved within ten minutes.•Acidic, acetonitrile rich mobile phase reduced hydrophobic interaction.•Electrostatic repulsion reduced the contribution of hydrophilic partition mechanism.•Sample diluent and mobile phase were optimized to avoid peak distortion by water.