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

Szymon Dziomba; Adrian Bekasiewicz; Adam Prahl; Tomasz Baczek; Piotr Kowalski

doi:10.1007/s00216-014-8104-1

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

Abstract

Derivatization is a frequently used sample prepara-tion procedure applicable to the enhancement of analyte de-tection sensitivity. Amino acids mostly require derivatization prior to electrophoretic or chromatographic analysis, especial-ly 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 pro-cess were discussed. The applicability of the elaborated meth-od was demons ated using human urine samples.

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Authors (5)

Szymon Dziomba
- Medical University of Gdańsk Department of Pharmaceutical Chemistry
Adrian Bekasiewicz dr hab. inż.
Adam Prahl
- Faculty of Chemistry, University of Gdańsk Institute of Organic Synthesis, Department of Organic Chemistry
Tomasz Baczek
- Medical University of Gdańsk Department of Pharmaceutical Chemistry
Piotr Kowalski
- Medical University of Gdańsk Department of Pharmaceutical Chemistry

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Category:: Articles
Type:: artykuł w czasopiśmie wyróżnionym w JCR
Published in:: ANALYTICAL AND BIOANALYTICAL CHEMISTRY no. 406, pages 6713 - 6721,
ISSN: 1618-2642
Language:: English
Publication year:: 2014
Bibliographic description:: Dziomba S., Bekasiewicz A., Prahl A., Baczek T., Kowalski P.: Improvement of derivatized amino acid detection sensitivity in micellar electrokinetic capillary chromatography by means of acid-induced pH-mediated stacking technique// ANALYTICAL AND BIOANALYTICAL CHEMISTRY. -Vol. 406, nr. 26 (2014), s.6713-6721
DOI:: Digital Object Identifier (open in new tab) 10.1007/s00216-014-8104-1
Verified by:: Gdańsk University of Technology