Document Detail


Determination of 13C isotopic enrichment of valine and threonine by GC-C-IRMS after formation of the N(O,S)-ethoxycarbonyl ethyl ester derivatives of the amino acids.
MedLine Citation:
PMID:  17468859     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We describe a new method of assessing, in a single run, (13)C isotopic enrichment of both Val and Thr by gas chromatography-combustion-isotope-ratio mass spectrometry (GC-C-IRMS). This method characterised by a rapid one-step derivatisation procedure performed at room temperature to form the N(O,S)-ethoxycarbonyl ethyl ester derivatives, and a polar column for GC. The suitability of this method for Val and Thr in in-vivo samples (mucosal hydrolysate) was demonstrated by studying protein metabolism with two tracers ((13)C-valine or (13)C-threonine). The intra-day and inter-day repeatability were both assessed either with standards or with in-vivo samples at natural abundance and at low (13)C isotopic enrichment. For inter-day repeatability CVs were between 0.8 and 1.5% at natural abundance and lower than 5.5% at 0.112 and 0.190 atom% enrichment for Val and Thr, respectively. Overall isotopic precision was studied for eleven standard amino acid derivatives (those of Val, Ala, Leu, Iso, Gly, Pro, Asp, Thr, Ser, Met, and Phe) and was assessed at 0.32 per thousand. The (13)C isotopic measurement was then extended to the other amino acids (Ala, Val, Leu, Iso, Gly, Pro, Thr, and Phe) at natural abundance for in-vivo samples. The isotopic precision was better than 0.002 atom% per amino acid (for n = 4 rats). This analytical method was finally applied to an animal study to measure Thr utilization in protein synthesis.
Authors:
Jean-Philippe Godin; Magali Faure; Denis Breuille; Gérard Hopfgartner; Laurent-Bernard Fay
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Publication Detail:
Type:  Journal Article     Date:  2007-04-28
Journal Detail:
Title:  Analytical and bioanalytical chemistry     Volume:  388     ISSN:  1618-2642     ISO Abbreviation:  Anal Bioanal Chem     Publication Date:  2007 Jun 
Date Detail:
Created Date:  2007-06-04     Completed Date:  2007-09-06     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101134327     Medline TA:  Anal Bioanal Chem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  909-18     Citation Subset:  IM    
Affiliation:
Department of Bioanalytical Science, Nestlé Research Center, Nestec Ltd, Vers chez les Blanc, Lausanne 26, Switzerland. jean-philippe.godin@rdls.nestle.com
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / chemistry*
Animals
Carbon Isotopes / chemistry*
Esters / chemistry*
Ethyl Ethers / chemistry*
Gas Chromatography-Mass Spectrometry / methods*
Intestinal Mucosa / chemistry,  metabolism
Male
Protein Biosynthesis
Rats
Rats, Sprague-Dawley
Reproducibility of Results
Sensitivity and Specificity
Threonine / analysis*
Time Factors
Valine / analysis*
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Carbon Isotopes; 0/Esters; 0/Ethyl Ethers; 7004-03-7/Valine; 72-19-5/Threonine

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


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