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Improvement of derivatized amino acid detection sensitivity in micellar electrokinetic capillary chromatography by means of acid-induced pH-mediated stacking technique

Derivatization is a frequently used sample preparation procedure applicable to the enhancement of analyte detection sensitivity. Amino acids mostly require derivatization prior to electrophoretic or chromatographic analysis, especially if spectrophotometric detection is used. This study presents an...

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Autores principales: Dziomba, Szymon, Bekasiewicz, Adrian, Prahl, Adam, Bączek, Tomasz, Kowalski, Piotr
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4182592/
https://www.ncbi.nlm.nih.gov/pubmed/25146356
http://dx.doi.org/10.1007/s00216-014-8104-1
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author Dziomba, Szymon
Bekasiewicz, Adrian
Prahl, Adam
Bączek, Tomasz
Kowalski, Piotr
author_facet Dziomba, Szymon
Bekasiewicz, Adrian
Prahl, Adam
Bączek, Tomasz
Kowalski, Piotr
author_sort Dziomba, Szymon
collection PubMed
description Derivatization is a frequently used sample preparation procedure applicable to the enhancement of analyte detection sensitivity. Amino acids mostly require derivatization prior to electrophoretic or chromatographic analysis, especially if spectrophotometric detection is used. This study presents an on-line preconcentration technique for derivatized amino acids. The sensitivity of the method was improved by the utilization of the proposed acid-induced pH-mediated stacking mechanism. The method is demonstrated by preconcentration of amino acids labeled with 2,4-dinitrofluorobenzene. Use of optimized conditions for a large sample volume injection (40 s, 13.8 kPa) followed by electrokinetic injection of 0.1 M HCl (20 s, 10 kV) gave a 20- to 30-fold enhancement of sensitivity. The significance of the sweeping mechanism and pseudo-isotachophoresis for the on-line sample focusing and the influence of parameters on the preconcentration process were discussed. The applicability of the elaborated method was demonstrated using human urine samples. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00216-014-8104-1) contains supplementary material, which is available to authorized users.
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spelling pubmed-41825922014-10-06 Improvement of derivatized amino acid detection sensitivity in micellar electrokinetic capillary chromatography by means of acid-induced pH-mediated stacking technique Dziomba, Szymon Bekasiewicz, Adrian Prahl, Adam Bączek, Tomasz Kowalski, Piotr Anal Bioanal Chem Research Paper Derivatization is a frequently used sample preparation procedure applicable to the enhancement of analyte detection sensitivity. Amino acids mostly require derivatization prior to electrophoretic or chromatographic analysis, especially if spectrophotometric detection is used. This study presents an on-line preconcentration technique for derivatized amino acids. The sensitivity of the method was improved by the utilization of the proposed acid-induced pH-mediated stacking mechanism. The method is demonstrated by preconcentration of amino acids labeled with 2,4-dinitrofluorobenzene. Use of optimized conditions for a large sample volume injection (40 s, 13.8 kPa) followed by electrokinetic injection of 0.1 M HCl (20 s, 10 kV) gave a 20- to 30-fold enhancement of sensitivity. The significance of the sweeping mechanism and pseudo-isotachophoresis for the on-line sample focusing and the influence of parameters on the preconcentration process were discussed. The applicability of the elaborated method was demonstrated using human urine samples. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00216-014-8104-1) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2014-08-22 2014 /pmc/articles/PMC4182592/ /pubmed/25146356 http://dx.doi.org/10.1007/s00216-014-8104-1 Text en © The Author(s) 2014 https://creativecommons.org/licenses/by/4.0/ Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.
spellingShingle Research Paper
Dziomba, Szymon
Bekasiewicz, Adrian
Prahl, Adam
Bączek, Tomasz
Kowalski, Piotr
Improvement of derivatized amino acid detection sensitivity in micellar electrokinetic capillary chromatography by means of acid-induced pH-mediated stacking technique
title Improvement of derivatized amino acid detection sensitivity in micellar electrokinetic capillary chromatography by means of acid-induced pH-mediated stacking technique
title_full Improvement of derivatized amino acid detection sensitivity in micellar electrokinetic capillary chromatography by means of acid-induced pH-mediated stacking technique
title_fullStr Improvement of derivatized amino acid detection sensitivity in micellar electrokinetic capillary chromatography by means of acid-induced pH-mediated stacking technique
title_full_unstemmed Improvement of derivatized amino acid detection sensitivity in micellar electrokinetic capillary chromatography by means of acid-induced pH-mediated stacking technique
title_short Improvement of derivatized amino acid detection sensitivity in micellar electrokinetic capillary chromatography by means of acid-induced pH-mediated stacking technique
title_sort improvement of derivatized amino acid detection sensitivity in micellar electrokinetic capillary chromatography by means of acid-induced ph-mediated stacking technique
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4182592/
https://www.ncbi.nlm.nih.gov/pubmed/25146356
http://dx.doi.org/10.1007/s00216-014-8104-1
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