Document Detail


Quantification and Mass Isotopomer Profiling of α-Keto Acids in Central Carbon Metabolism.
MedLine Citation:
PMID:  24533614     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Mass spectrometry has been established as a powerful and versatile technique for studying cellular metabolism. Applications range from profiling of metabolites to accurate quantification and tracing of stable isotopes through the biochemical reaction network. Despite broad coverage of central carbon metabolism, most methods fail to provide accurate assessments of the α-keto acids oxaloacetic acid, pyruvate, and glyoxylate because these compounds are highly reactive and degraded during sample processing and mass spectrometric measurement. We present a derivatization procedure to chemically stabilize these compounds readily during quenching of cellular metabolism. Stable derivatives were analyzed by ultrahigh pressure liquid chromatography coupled tandem mass spectrometry to accurately quantify the abundance of α-keto acids in biological matrices. Eventually, we demonstrated that the developed protocol is suited to measure mass isotopomers of these α-keto acids in tracer studies with stable isotopes. In conclusion, the here described method fills one of the last technical gaps for metabolomics investigations of central carbon metabolism.
Authors:
Michael Zimmermann; Uwe Sauer; Nicola Zamboni
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Publication Detail:
Type:  Journal Article     Date:  2014-02-24
Journal Detail:
Title:  Analytical chemistry     Volume:  86     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2014 Mar 
Date Detail:
Created Date:  2014-03-18     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3232-7     Citation Subset:  IM    
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