Document Detail


Fluorescein thiocarbamyl amino acids as internal standards for migration time correction in capillary sieving electrophoresis.
MedLine Citation:
PMID:  19249052     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A number of algorithms have been developed to correct for migration time drift in capillary electrophoresis. Those algorithms require identification of common components in each run. However, not all components may be present or resolved in separations of complex samples, which can confound attempts for alignment. This paper reports the use of fluorescein thiocarbamyl derivatives of amino acids as internal standards for alignment of 3-(2-furoyl)quinoline-2-carboxaldehyde (FQ)-labeled proteins in capillary sieving electrophoresis. The fluorescein thiocarbamyl derivative of aspartic acid migrates before FQ-labeled proteins and the fluorescein thiocarbamyl derivative of arginine migrates after the FQ-labeled proteins. These compounds were used as internal standards to correct for variations in migration time over a two-week period in the separation of a cellular homogenate. The experimental conditions were deliberately manipulated by varying electric field and sample preparation conditions. Three components of the homogenate were used to evaluate the alignment efficiency. Before alignment, the average relative standard deviation in migration time for these components was 13.3%. After alignment, the average relative standard deviation in migration time for these components was reduced to 0.5%.
Authors:
Haley R Pugsley; Kristian E Swearingen; Norman J Dovichi
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2009-02-10
Journal Detail:
Title:  Journal of chromatography. A     Volume:  1216     ISSN:  1873-3778     ISO Abbreviation:  J Chromatogr A     Publication Date:  2009 Apr 
Date Detail:
Created Date:  2009-03-23     Completed Date:  2009-09-10     Revised Date:  2011-05-09    
Medline Journal Info:
Nlm Unique ID:  9318488     Medline TA:  J Chromatogr A     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  3418-20     Citation Subset:  IM    
Affiliation:
Department of Chemistry, University of Washington, Seattle, WA 98195-1700, USA.
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MeSH Terms
Descriptor/Qualifier:
Algorithms
Arginine* / analogs & derivatives,  chemistry
Aspartic Acid* / analogs & derivatives,  chemistry
Cell Line, Tumor
Electrophoresis, Capillary / methods*
Female
Fluorescein / chemistry*
Furans / analysis
Humans
Quinolines / analysis
Reference Standards
Grant Support
ID/Acronym/Agency:
R01 NS061767-02/NS/NINDS NIH HHS; R01NS061767/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Furans; 0/Quinolines; 126769-01-5/3-(2-furoyl)quinoline-2-carbaldehyde; 2321-07-5/Fluorescein; 56-84-8/Aspartic Acid; 74-79-3/Arginine
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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