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  • On-line sample preconcentration by sweeping and poly(ethylene oxide)-mediated stacking for simultaneous analysis of nine pairs of amino acid enantiomers in capillary electrophoresis.

On-line sample preconcentration by sweeping and poly(ethylene oxide)-mediated stacking for simultaneous analysis of nine pairs of amino acid enantiomers in capillary electrophoresis.

Talanta (2013-08-21)
En-Ping Lin, Kai-Cheng Lin, Chia-Wei Chang, Ming-Mu Hsieh
ZUSAMMENFASSUNG

This study proposes a sensitive method for the simultaneous separation and concentration of 9 pairs of amino acid enantiomers by combining poly(ethylene oxide) (PEO)-based stacking, β-cyclodextrin (β-CD)-mediated micellar electrokinetic chromatography (MEKC), and 9-fluoroenylmethyl chloroformate (FMOC) derivatization. The 9 pairs of FMOC-derivatized amino acid enantiomers were baseline separated using a discontinuous system, and the buffer vials contained a solution of 150 mM Tris-borate (TB), 12.5% (v/v) isopropanol (IPA), 0.5% (w/v) PEO, 35 mM sodium taurodeoxycholate (STDC), and 35 mM β-CD, and the capillary was filled with a solution of 1.5 M TB, 12.5% (v/v) IPA, 35 mM STDC, and 35 mM β-CD. Based on the difference in viscosity between the sample zone and PEO solution and because of the STDC sweeping, the discontinuous system effectively stacked 670 nL of the 9 pairs of FMOC-derivatized amino acid enantiomers without losing chiral resolution. Consequently, the limits of detection for the 9 pairs of FMOC-derivatized amino acid enantiomers were reduced to 40-60 nM. This method was successfully used to determine d-Tryptophan (Trp), l-Trp, d-Phenylalanine (Phe), l-Phe, d-Glutamic acid (Glu), and l-Glu in various types of beers.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Phthaldialdehyd, ≥97% (HPLC), powder or crystals
Sigma-Aldrich
β-Cyclodextrin, ≥97%
Sigma-Aldrich
Fmoc-Chlorid, 97%
Supelco
Fmoc-Chlorid, derivatization grade (HPLC), LiChropur, ≥99.0% (HPLC)
Sigma-Aldrich
Phthaldialdehyd, suitable for fluorescence, ≥99.0% (HPLC)
Sigma-Aldrich
Phthaldialdehyde Reagent, Solution Complete
Sigma-Aldrich
Phthaldialdehyd, suitable for HPLC fluorimetric detection of amino acids, ≥99% (HPLC), powder or crystals
Sigma-Aldrich
β-Cyclodextrin, powder, BioReagent, suitable for cell culture, ≥97%
Sigma-Aldrich
2,3-Naphthalindicarboxaldehyd, suitable for fluorescence
Sigma-Aldrich
Fmoc-Chlorid, BioReagent, ≥99.0% (HPLC)
Sigma-Aldrich
β-Cyclodextrin, Wacker Chemie AG, ≥95.0% (HPLC)
Sigma-Aldrich
β-Cyclodextrin, Wacker Chemie AG
Sigma-Aldrich
β-Cyclodextrin, Wacker Chemie AG, ≥98.0% (HPLC)
Sigma-Aldrich
Phthaldialdehyde Reagent, Solution Incomplete