Liquid chromatographic enantiomer separation of α-amino acids and their methyl and ethyl esters as fluorenylmethoxycarbonyl (FMOC) derivatives was performed using a recently developed chiral column (Chiralpak IE) based on polysaccharide derivative under simultaneous UV detection and fluorescence detection. The degree of enantiomer separation of α-amino acid esters as FMOC derivatives is generally higher than that of the corresponding α-amino acids. Especially, α-amino acid methyl esters showed the greatest enantioseparation. As this method developed in this study can be applied to determine the chemical and optical purity of α-amino acids and esters, it is expected to be quite useful for their chiral separation using Chiralpak IE.