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Since most of the proteinogenic alpha-amino acids contain a chiral carbon atom, the stereoisomers of both these amino acids and the peptides in which they are to be found may possess differences in biological activity in living systems. The important analytical task of the separation of optical isomers is achieved mainly by chromatographic methods. This special review surveys direct and indirect HPLC separations of biologically important, small epimeric peptides and their L, D-amino acid content.