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Stacking and separation of fluorescent derivatives of amino acids by micellar electrokinetic chromatography in the presence of poly(ethylene oxide).
J Chromatogr A. 2007 Mar 30; 1146(1):118-24.JC

Abstract

A new approach for the analysis of large-volume naphthalene-2,3-dicarboxaldehyde (NDA) derivatives of amino acids by micellar electrokinetic chromatography (MEKC) in conjunction with a purple light-emitting diode-induced fluorescence detection is described. In order to optimize resolution, speed, and stacking efficiency, a discontinuous condition is essential for the analysis of NDA-amino acid derivatives. The optimum conditions use 2.0M TB (pH 10.0) buffer containing 40mM sodium dodecyl sulfate (SDS) to fill the capillary, deionized water to dilute samples, and 200mM TB (pH 9.0) containing 10mM SDS to prepare 0.6% poly(ethylene oxide) (PEO). Once high voltage is applied, PEO solution enters the capillary via electroosmotic flow and SDS micelles interact and thus sweep the NDA-amino acid derivatives having smaller electrophoretic mobilities than that of SDS micelles in the sample zone. When the aggregates between SDS micelles and NDA amino acid derivatives enter PEO zone, they are stacked due to decrease in electric field and increases in viscosity. Under the optimum conditions, the concentration and separation of 0.53-microL 13 NDA-amino acid derivatives that are negatively charged has been demonstrated by using a 60-cm capillary, with the efficiencies 0.3-9.0x10(5) theoretical plates and the LODs at signal-to-noise ratio 3 ranging from 0.30 to 2.76nM. When compared to standard injection (30-cm height for 10s), the approach allows the sensitivity enhancements over the range of 50-800 folds for the derivatives. The new approach has been applied to the analysis of a red wine sample, with great linearity of fluorescent intensity against concentrations (R(2)>0.98) and the RSD (three repetitive runs in one day) values of the migration times for the ten identified amino acids less than 2.8%.

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Publisher Full Text

Authors+Show Affiliations

Chiu TC
Department of Chemistry, National Taiwan University, Taipei, Taiwan.
Chang HT
No affiliation info available

MeSH

Amino AcidsChromatography, Micellar Electrokinetic CapillaryFluorescenceNaphthalenesPolyethylene GlycolsReproducibility of ResultsSodium Dodecyl SulfateWine

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17300792

Citation

Chiu, Tai-Chia, and Huan-Tsung Chang. "Stacking and Separation of Fluorescent Derivatives of Amino Acids By Micellar Electrokinetic Chromatography in the Presence of Poly(ethylene Oxide)." Journal of Chromatography. A, vol. 1146, no. 1, 2007, pp. 118-24.
Chiu TC, Chang HT. Stacking and separation of fluorescent derivatives of amino acids by micellar electrokinetic chromatography in the presence of poly(ethylene oxide). J Chromatogr A. 2007;1146(1):118-24.
Chiu, T. C., & Chang, H. T. (2007). Stacking and separation of fluorescent derivatives of amino acids by micellar electrokinetic chromatography in the presence of poly(ethylene oxide). Journal of Chromatography. A, 1146(1), 118-24.
Chiu TC, Chang HT. Stacking and Separation of Fluorescent Derivatives of Amino Acids By Micellar Electrokinetic Chromatography in the Presence of Poly(ethylene Oxide). J Chromatogr A. 2007 Mar 30;1146(1):118-24. PubMed PMID: 17300792.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Stacking and separation of fluorescent derivatives of amino acids by micellar electrokinetic chromatography in the presence of poly(ethylene oxide). AU - Chiu,Tai-Chia, AU - Chang,Huan-Tsung, Y1 - 2007/02/04/ PY - 2006/08/25/received PY - 2007/01/20/revised PY - 2007/01/26/accepted PY - 2007/2/16/pubmed PY - 2007/5/1/medline PY - 2007/2/16/entrez SP - 118 EP - 24 JF - Journal of chromatography. A JO - J Chromatogr A VL - 1146 IS - 1 N2 - A new approach for the analysis of large-volume naphthalene-2,3-dicarboxaldehyde (NDA) derivatives of amino acids by micellar electrokinetic chromatography (MEKC) in conjunction with a purple light-emitting diode-induced fluorescence detection is described. In order to optimize resolution, speed, and stacking efficiency, a discontinuous condition is essential for the analysis of NDA-amino acid derivatives. The optimum conditions use 2.0M TB (pH 10.0) buffer containing 40mM sodium dodecyl sulfate (SDS) to fill the capillary, deionized water to dilute samples, and 200mM TB (pH 9.0) containing 10mM SDS to prepare 0.6% poly(ethylene oxide) (PEO). Once high voltage is applied, PEO solution enters the capillary via electroosmotic flow and SDS micelles interact and thus sweep the NDA-amino acid derivatives having smaller electrophoretic mobilities than that of SDS micelles in the sample zone. When the aggregates between SDS micelles and NDA amino acid derivatives enter PEO zone, they are stacked due to decrease in electric field and increases in viscosity. Under the optimum conditions, the concentration and separation of 0.53-microL 13 NDA-amino acid derivatives that are negatively charged has been demonstrated by using a 60-cm capillary, with the efficiencies 0.3-9.0x10(5) theoretical plates and the LODs at signal-to-noise ratio 3 ranging from 0.30 to 2.76nM. When compared to standard injection (30-cm height for 10s), the approach allows the sensitivity enhancements over the range of 50-800 folds for the derivatives. The new approach has been applied to the analysis of a red wine sample, with great linearity of fluorescent intensity against concentrations (R(2)>0.98) and the RSD (three repetitive runs in one day) values of the migration times for the ten identified amino acids less than 2.8%. SN - 0021-9673 UR - https://www.unboundmedicine.com/medline/citation/17300792/Stacking_and_separation_of_fluorescent_derivatives_of_amino_acids_by_micellar_electrokinetic_chromatography_in_the_presence_of_poly_ethylene_oxide__ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9673(07)00219-1 DB - PRIME DP - Unbound Medicine ER -