ABSTRACT
A reversed phase high performance liquid chromatographic ( RP-HPLC ) method was proposed by using C18 column tandem crown ether chiral column for simultaneous separation of three kinds of aromatic amino acid enantiomers. The chromatographic conditions, including mobile phase proportion, pH, column temperature and flow rate, were optimized. The experimental results showed that the three aromatic amino acid enantiomers could be successfully separated under the optimal conditions such as perchloric acid solution-acetonitrile ( 86:14, V/V, pH = 2 ) as mobile phase, column temperature of 20℃ and flow rate of 0. 4 mL/min. The separation under four HPLC column connection modes was further investigated. The results revealed that different kinds of amino acids could be separated on C18 column, but not for amino acid enantiomers; amino acid enantiomers could be separated on crown ether chiral column, but the chromatographic peaks of each amino acid enantiomers overlapped seriously; baseline separation of three amino acid enantiomers could be obtained under the connection of two columns, but the separation under different column connection sequence almost had no difference except the peak shape.