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Title: Indirect fluorescence detection of native amino acids in capillary zone electrophoresis

Abstract

Amino acids are but one of several important classes of small chemical compounds in biological chemistry that have an inherent lack of analytically useful physical properties. Amino acids, peptides, fatty acids, sugars, many mono-, di-, and tricarboxylic acids, and phosphorylated intermediates in glycolysis and metabolism show little, if any, UV or visible absorption, fluorescence, or electrochemical activity. As the emphasis of biochemical research shifts to smaller samples where, for example, picomolar quantities of amino acids are analyzed in gas phase protein sequencing or in microliter samples of the extracellular fluid of the mammalian brain, the analytical problem becomes even more challenging due to the small volume of sample available for analysis. In this work, laser-induced fluorescence spectroscopy is performed on-column to detect the bands separated with capillary zone electrophoresis (CZE). CZE is an instrumental form of zone electrophoresis where chemical species are separated purely on the basis of their electrophoretic mobility, since no supporting gel is utilized. Both anions and cations can be separated in the same run because of the large electroosmotic flow generated in small diameter capillaries. This technique has already been used successfully in the rapid, efficient separation of dansyl-amino acids.

Authors:
;
Publication Date:
Research Org.:
Iowa State Univ., Ames (USA)
OSTI Identifier:
7014033
DOE Contract Number:  
W-7405-ENG-82
Resource Type:
Journal Article
Journal Name:
Anal. Chem.; (United States)
Additional Journal Information:
Journal Volume: 60:17
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY; AMINO ACIDS; FLUORESCENCE SPECTROSCOPY; ELECTROPHORESIS; EXPERIMENTAL DATA; LASERS; QUANTITATIVE CHEMICAL ANALYSIS; CARBOXYLIC ACIDS; CHEMICAL ANALYSIS; DATA; EMISSION SPECTROSCOPY; INFORMATION; NUMERICAL DATA; ORGANIC ACIDS; ORGANIC COMPOUNDS; SPECTROSCOPY; 400102* - Chemical & Spectral Procedures; 400105 - Separation Procedures

Citation Formats

Kuhr, W G, and Yeung, E S. Indirect fluorescence detection of native amino acids in capillary zone electrophoresis. United States: N. p., 1988. Web. doi:10.1021/ac00168a038.
Kuhr, W G, & Yeung, E S. Indirect fluorescence detection of native amino acids in capillary zone electrophoresis. United States. doi:10.1021/ac00168a038.
Kuhr, W G, and Yeung, E S. Thu . "Indirect fluorescence detection of native amino acids in capillary zone electrophoresis". United States. doi:10.1021/ac00168a038.
@article{osti_7014033,
title = {Indirect fluorescence detection of native amino acids in capillary zone electrophoresis},
author = {Kuhr, W G and Yeung, E S},
abstractNote = {Amino acids are but one of several important classes of small chemical compounds in biological chemistry that have an inherent lack of analytically useful physical properties. Amino acids, peptides, fatty acids, sugars, many mono-, di-, and tricarboxylic acids, and phosphorylated intermediates in glycolysis and metabolism show little, if any, UV or visible absorption, fluorescence, or electrochemical activity. As the emphasis of biochemical research shifts to smaller samples where, for example, picomolar quantities of amino acids are analyzed in gas phase protein sequencing or in microliter samples of the extracellular fluid of the mammalian brain, the analytical problem becomes even more challenging due to the small volume of sample available for analysis. In this work, laser-induced fluorescence spectroscopy is performed on-column to detect the bands separated with capillary zone electrophoresis (CZE). CZE is an instrumental form of zone electrophoresis where chemical species are separated purely on the basis of their electrophoretic mobility, since no supporting gel is utilized. Both anions and cations can be separated in the same run because of the large electroosmotic flow generated in small diameter capillaries. This technique has already been used successfully in the rapid, efficient separation of dansyl-amino acids.},
doi = {10.1021/ac00168a038},
journal = {Anal. Chem.; (United States)},
number = ,
volume = 60:17,
place = {United States},
year = {1988},
month = {9}
}