Assay of the concentration and 13C enrichment of acetate and acetyl-CoA by gas chromatography-mass spectrometry.
We present two techniques for determining the concentration and 13C enrichment of acetate in biological fluids. After the sample has been spiked with an internal standard of [2,2,2,2H3,1-13C]acetate, acetate is first enzymatically converted to acetyl-coenzyme A, which is chemically converted to acetylglycine. The latter is analyzed by gas chromatography-mass spectrometry, either as a methyl ester by positive chemical ionization or as a pentafluorobenzyl ester by negative chemical ionization. The mole percentage enrichment of tissue acetyl-CoA can also be assayed after conversion to acetylglycine pentafluorobenzyl ester.
David
F
F
Department of Nutrition, Case Western Reserve University, Cleveland, Ohio 44106.
Beylot
M
M
Reider
M W
MW
Anderson
V E
VE
Brunengraber
H
H
D000085:Acetates / Q000032:analysis* / Q000737:chemistry; D000105:Acetyl Coenzyme A / Q000032:analysis* / Q000737:chemistry; D000818:Animals; D002247:Carbon Isotopes; D004285:Dogs; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry; D016014:Linear Models; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley
D000085:Acetates; D002247:Carbon Isotopes; D000105:Acetyl Coenzyme A