Indirect fluorescence detection of native amino acids in capillary zone electrophoresis
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.
- Research Organization:
- Iowa State Univ., Ames (USA)
- DOE Contract Number:
- W-7405-ENG-82
- OSTI ID:
- 7014033
- Journal Information:
- Anal. Chem.; (United States), Vol. 60:17
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
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