Document Detail

A comparison of fluorescamine and naphthalene-2,3-dicarboxaldehyde fluorogenic reagents for microplate-based detection of amino acids.
MedLine Citation:
PMID:  11673879     Owner:  NLM     Status:  MEDLINE    
The use of appropriate fluorometric derivatization procedures is of considerable importance for accurate determination of amino acids in biological samples and in metal-assisted peptide hydrolysis reactions. It is especially critical for the relative fluorescence intensities (RFI) of equal amounts of amino acids to be as similar as possible. While fluorescamine and naphthalene-2,3-dicarboxaldehyde (NDA) have proven to be excellent fluorogenic reagents for amino acid detection, the effects of various factors such as organic solvent, buffer, and pH have never been rigorously evaluated with respect to normalizing the relative fluorescence intensities of individual amino acids. To this end, here we describe optimized fluorescamine and NDA derivatization reactions that enhance the accuracy of microplate-based detection of amino acids. For both fluorescamine and NDA, we have shown that the RFI values of 16 of 19 amino acids are greater than 70%. Although determination of tryptophan is problematic, this difficulty is overcome by the addition of beta-cyclodextrin to the NDA reaction. In principle, the optimized fluorescamine and NDA microplate procedures reported here can be utilized as complementary techniques for the detection of 19 of 20 naturally occurring amino acids.
T Bantan-Polak; M Kassai; K B Grant
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Analytical biochemistry     Volume:  297     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2001 Oct 
Date Detail:
Created Date:  2001-10-24     Completed Date:  2002-01-18     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  128-36     Citation Subset:  IM    
Copyright Information:
Copyright 2001 Academic Press.
Department of Chemistry, Center for Biotechnology and Drug Design, Georgia State University, University Plaza, Atlanta, Georgia 30303-3083, USA.
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MeSH Terms
Amino Acids / analysis*
Cyclodextrins / chemistry*
Dipeptides / analysis
Fluorescamine / chemistry*
Fluorescent Dyes / chemistry
Fluorometry / methods*
Naphthalenes / chemistry*
Platinum / chemistry
Reproducibility of Results
Tryptophan / analysis*
Reg. No./Substance:
0/Amino Acids; 0/Cyclodextrins; 0/Dipeptides; 0/Fluorescent Dyes; 0/Naphthalenes; 0/beta-Cyclodextrins; 38183-12-9/Fluorescamine; 7149-49-7/2,3-naphthalenedicarboxaldehyde; 73-22-3/Tryptophan; 7440-06-4/Platinum; 7585-39-9/betadex

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

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