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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
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
β-Cyclodextrin, Produced by Wacker Chemie AG, Burghausen, Germany, Life Science, ≥98.0% (HPLC)
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Sigma-Aldrich
β-Cyclodextrin, produced by Wacker Chemie AG, Burghausen, Germany, ≥95.0% (HPLC)
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Sigma-Aldrich
Phthaldialdehyde, suitable for HPLC fluorimetric detection of amino acids, ≥99% (HPLC), powder or crystals
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Sigma-Aldrich
β-Cyclodextrin, produced by Wacker Chemie AG, Burghausen, Germany
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Sigma-Aldrich
Phthaldialdehyde, for fluorescence, ≥99.0% (HPLC)
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Sigma-Aldrich
Phthaldialdehyde, ≥97% (HPLC), powder or crystals
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Sigma-Aldrich
β-Cyclodextrin, powder, BioReagent, suitable for cell culture, ≥97%
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Sigma-Aldrich
β-Cyclodextrin, ≥97%
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Sigma-Aldrich
2,3-Naphthalenedicarboxaldehyde, suitable for fluorescence
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Phthaldialdehyde Reagent, Solution Incomplete
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Sigma-Aldrich
Phthaldialdehyde Reagent, Solution Complete
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Supelco
Fmoc chloride, for HPLC derivatization, LiChropur, ≥99.0% (HPLC)
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Sigma-Aldrich
Fmoc chloride, 97%
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Sigma-Aldrich
Fmoc chloride, BioReagent, ≥99.0% (HPLC)
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