Document Detail


Fluorimetric determination of intra- and extracellular free amino acids in the microalgae Tetraselmis gracilis (Prasinophyceae) using monolithic column in reversed phase mode.
MedLine Citation:
PMID:  19569093     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This paper describes the development and application of an RP HPLC method using a C(18) monolithic stationary phase for the separation and quantification of extra- and intracellular amino acids in a batch cultivation of the marine alga Tetraselmis gracilis. Fluorimetric detection was made after separation of the o-phthaldialdehyde 2-mercaptoethanol (OPA-2MCE) derivatives using a binary gradient elution. Separation of 19 amino acids was achieved with resolution >1.5 in about 39 min at a flow rate of 1.5 mL/min. RSD of analyses in seawater medium ranged from 0.36% for Orn (0.50 micromol/L) to 12% for Ile (0.10 micromol/L). The main constituents of the intracellular dissolved free amino acids (DFAAs) in the exponential growth phase were arginine (Arg), asparagine (Asn), alanine (Ala), aspartic acid (Asp), glutamic acid (Glu), serine (Ser), glycine (Gly), glutamine (Gln), and leucine (Leu). The major amino acids excreted to the media were valine (Val), Ala, Ser, and Gly. The monolithic phase facilitates the analysis by shortening the separation time and saving solvents and instrumentation costs (indeed conventional HPLC instrumentation can be used, running at lower pressures than those ones used with packed particle columns).
Authors:
José Carlo Pires Penteado; Marilda Rigobello-Masini; Cleber Wanderlei Liria; M Terêsa Machini Miranda; Jorge Cesar Masini
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of separation science     Volume:  32     ISSN:  1615-9314     ISO Abbreviation:  J Sep Sci     Publication Date:  2009 Aug 
Date Detail:
Created Date:  2009-08-17     Completed Date:  2009-10-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101088554     Medline TA:  J Sep Sci     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  2827-34     Citation Subset:  IM    
Affiliation:
Departamento de Química Fundamental, Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brazil.
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MeSH Terms
Descriptor/Qualifier:
Algae, Green* / chemistry,  genetics
Amino Acids / analysis*
Anion Exchange Resins / chemistry
Chromatography, High Pressure Liquid
Chromatography, Ion Exchange
Fluorometry / instrumentation,  methods*
Seawater
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Anion Exchange Resins

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


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