Journal
ELECTROPHORESIS
Volume 30, Issue 16, Pages 2837-2844Publisher
WILEY
DOI: 10.1002/elps.200800799
Keywords
CE-ESI-MS; Chiral peptide; Crown ethers; Enantiomeric separation; Partial filling CE
Funding
- National Nature Sciences Foundation of China [20675016, 20735002, 20775014]
- Key Program of Science and Technology Department of Fujian Province, China [200710020]
- Key Special Purpose Foundation of Physical Education Bureau of Fujian Province [HX2005-74]
- Plan of the University New Century Outstanding Talent of Fujian Province [HX2006-101]
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Enantiomer of chiral dipeptides were separated by CE-ESI-MS in a bare fused-silica capillary using (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid (18C(6)H(4)) as the chiral selector. As 18C(6)H(4) is a kind of nonvolatile chiral selector, in order to prevent from 18C(6)H(4) into the ion-source of CE-ESI-MS, a partial filling technique was employed in this study. Some dipeptides with one chiral center or two chiral centers, such as DL-Leu-DL-Leu, D-Ala-D-Ala and L-Ala-L-Ala, Gly-D-Phe and Gly-L-Phe were used to evaluate this CE-ESI-MS system. Optimized conditions were achivevd with 2.0 mol/L acetic acid (pH 2.15) as the running electrolyte, 5 mM 18C(6)H(4) in 3.0 mol/L acetic acid (pH 2.00) was injected hydrodynamically (50 mbar for 960 s) before sample injection. In total 7.5 mM acetic acid in 80% v/v methanol-water was used as the sheath liquid, and 20 kV applied voltage was used. Under the optimum conditions, these dipeptides were separated and detected. LODs (defined as S/N = 3) of this method were 0.20, 0.10, 0.05 and 0.10 mu mol/L for D-Ala-D-Ala, L-Ala-L-Ala, DL-LeU-DL-Leu, Gly-L-Phe and Gly-D-Phe, respectively. The RSDs (n = 7) of the method were 0.68-2.08% for migration times and 2.32-5.24% for peak areas. The proposed method was also successfully applied to the enantioselective analysis of these dipeptides in the spiked serum samples with satisfactory results.
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