Abstract
A new capillary electrophoresis method was applied to chiral separation of three amino acids, including D,L-tryptophan, D,L-tyrosine and D,L-phenylalanine. The chiral resolution was attained in an untreated fused-sillica capillary using a dual chiral selector, which was made up of Cu(II)-L-histidine complex and β-cyclodextrin (CD). The cardinal factors influencing its separation efficiency, such as chiral selectors, buffer pH and applied voltage, were optimized. Best results were acquired by using a buffer consisting of 10 mmol/L Cu(II), 13 mmol/L L-histidine, 8 mmol/L β-CD, 5 mmol/L phosphate adjusted to pH 5.0 and 15 kV applied voltage. All enantiomers were entirely resolved within 20 min with high resolutions of 3.6~6.1. The analysis method was verified through the determination of D,L-tryptophan in terms of linearity, precision and accuracy. And the robustness of this method was proved. The Limit of Detection and Limit of Quantification for both enantiomers were 2.5 and 5 μg/mL, respectively. The method was perfectly applied to the determination of the enantiomeric purity of L-tryptophan. Furthermore, the interaction between Cu(II)-L-histidine complex and β-CD was also studied using Ultraviolet-visible and 1H NMR spectroscopy to explain the synergistic effect involved. The results illustrated that Cu(II)-L-histidine complex and β-CD played a synergistic role in the enantiomeric separation of chiral drugs, with good prospects for application.
Recent Advances in SPE-Chiral-HPLC Methods for Enantiomeric Separation of Chiral Drugs in Biological Samples
