| Capillary electrophoresis of free amino acids in physiological fluids without derivatization employing direct or indirect absorbance detection. | |
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MedLine Citation:
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PMID: 22125149 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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Whole blood and/or plasma amino acids are useful for monitoring whole body protein and amino acid metabolism in an organism under various physiological and pathophysiological conditions. Various methodological procedures are in use for their measurement in biological fluids. From the time when capillary electrophoresis was introduced as a technology offering rapid separation of various ionic and/or ionizable compounds with low sample and solvent consumption, there were many attempts to use it for the measurement of amino acids present in physiological fluids. As a rule, these methods require derivatization procedures for detection purposes.Here, we present two protocols for the analysis of free amino acids employing free zone capillary electrophoresis. Main advantage of both methods is an absence of any derivatization procedures that permits the analysis of free amino acid in physiological fluids. The method using direct detection and carrier electrolyte consisting of disodium monophosphate (10 mM at pH 2.90) permits determination of compounds that absorb in UV region (aromatic and sulfur containing amino acids, as well as some peptides, such as carnosine, reduced and oxidized glutathione). The other method uses indirect absorbance detection, employing 8 mM p-amino salicylic acid buffered with sodium carbonate at pH 10.2 as running electrolyte. It permits quantification of 30 underivatized physiological amino acids and peptides. In our experience, factorial design represents a useful tool for final optimization of the electrophoretic conditions if it is necessary. |
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Authors:
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Gordana D Zunić; Slavica Spasić; Zorana Jelić-Ivanović |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 828 ISSN: 1940-6029 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2012 |
Date Detail:
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Created Date: 2011-11-29 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 243-54 Citation Subset: IM |
Affiliation:
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Institute for Medical Research, Military Medical Academy, Belgrade, Serbia, gordana144@open.telekom.rs. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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