An isotope-dilution, GC–MS assay for formate and its application to human and animal metabolism

Formate, a crucial component of one-carbon metabolism, is increasingly recognized as an important intermediate in production and transport of one-carbon units. Unlike tetrahydrofolate-linked intermediates, it is not restricted to the intracellular milieu so that circulating levels of formate can pro...

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Bibliographic Details
Published in:Amino acids 2014-08, Vol.46 (8), p.1885-1891
Main Authors: Lamarre, Simon G., MacMillan, Luke, Morrow, Gregory P., Randell, Edward, Pongnopparat, Theerawat, Brosnan, Margaret E., Brosnan, John T.
Format: Article
Language:English
Subjects:
Amino acids
Analytical Chemistry
Animals
Assaying
Biochemical Engineering
Biochemistry
Biomedical and Life Sciences
Cellular
Circulating
Female
Fluorobenzenes - chemistry
Formates
Formates - analysis
Formates - blood
Formates - urine
Gas Chromatography-Mass Spectrometry - methods
Human
Humans
Life Sciences
Male
Metabolism
Neurobiology
Original Article
Proteomics
Rats
Rats, Sprague-Dawley
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Summary:Formate, a crucial component of one-carbon metabolism, is increasingly recognized as an important intermediate in production and transport of one-carbon units. Unlike tetrahydrofolate-linked intermediates, it is not restricted to the intracellular milieu so that circulating levels of formate can provide insight into cellular events. We report a novel isotope-dilution, GC–MS assay employing derivatization by 2,3,4,5,6-pentafluorobenzyl bromide for the determination of formate in biological samples. This assay is robust and sensitive; it may be applied to the measurement of formate in serum, plasma and urine. We demonstrate how this method may be applied by providing the first characterization of formate levels in a human population; formate levels were higher in males than in females. We also show how this procedure may be applied for the measurement of in vivo kinetics of endogenous formate production in experimental animals.
ISSN:0939-4451
1438-2199
DOI:10.1007/s00726-014-1738-7