The resolution of amino acid enantiomers is important in peptide synthesis and structure determinations. Chromatographic methods of analysis have the advantage of resolving not only a pair of enantiomers, but also a mixture of several amino acid enantiomers. Liquid chromatographic resolution of optical isomers is based either on the formation of diastereomers or on the application of chiral stationary or mobile phases. The separation of racemates into enantiomers by means of column liquid chromatography is a rapidly developing field covering theoretical studies on the underlying retention mechanism and practical studies. The retention of amino acids in the normal-phase system differed considerably from that in the reversed-phase system, hydrophobic amino acids eluting first followed by hydrophilic amino acids. The use of dansyl derivatives rather than the free amino acids allows their chromatographic behavior to be studied by monitoring the fluorescence.