title: Assay of the concentration and 13C enrichment of acetate and acetyl-CoA by gas chromatography-mass spectrometry.
abstract: 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.
authors.affiliation: Department of Nutrition, Case Western Reserve University, Cleveland, Ohio 44106.
authors.forename: F
authors.initials: F
authors.lastname: David
authors.forename: M
authors.initials: M
authors.lastname: Beylot
authors.forename: M W
authors.initials: MW
authors.lastname: Reider
authors.forename: V E
authors.initials: VE
authors.lastname: Anderson
authors.forename: H
authors.initials: H
authors.lastname: Brunengraber
mesh_terms: 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
chemical_list: D000085:Acetates; D002247:Carbon Isotopes; D000105:Acetyl Coenzyme A