Document Detail


Sequential injection chromatography for fluorimetric determination of intracellular amino acids in marine microalgae.
MedLine Citation:
PMID:  22125154     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
This chapter describes a sequential injection chromatography method to automate the fluorimetric determination of amino acids after precolumn derivatization with o-phthaldialdehyde in presence of 2-mercaptoethanol using reverse-phase liquid chromatography in C(18) silica-based monolithic column. The method is low-priced and based on six steps of isocratic elutions. At a flow rate of 30 μl/s, a 25 mm long-column coupled to 5-mm guard column is capable to separate aspartic acid (Asp), glutamic acid (Glu), asparagine (Asn), serine (Ser), glycine (Gly), threonine (Thr), citrulline (Ctr), arginine (Arg), alanine (Ala), tyrosine (Tyr), phenylalanine (Phe), ornithine (Orn), and lysine (Lys). Under these conditions, histidine (His) and glutamine (Gln), methionine (Met) and valine (Val), and isoleucine (Ile) and leucine (Leu) coelute. The entire cycle of amino acids derivatization, chromatographic separation, and column conditioning at the end of separation lasts 16 min. The method was successfully applied to the determination of the major intracellular free amino acids in the marine green alga Tetraselmis gracilis.
Authors:
Marilda Rigobello-Masini; Jorge C Masini
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  828     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2012  
Date Detail:
Created Date:  2011-11-29     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  305-15     Citation Subset:  IM    
Affiliation:
Instituto de Química, Universidade de São Paulo, São Paulo, Brazil.
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