Gas chromatography-mass spectrometry assay of the (18)o enrichment of water as trimethyl phosphate.
We have developed an assay for determining the 18O enrichment of water in biological fluids. Urine, plasma, or whole blood is reacted with phosphorous pentachloride to yield phosphoric acid. Derivatization of phosphoric acid with diazomethane generates trimethyl phosphate. The enrichment of trimethyl phosphate is nearly four times that of water and is assayed using gas chromatography-mass spectrometry (electron impact ionization). Yang et al. (1998, Anal. Biochem. 258, 315-321) assayed the 2H enrichment of body water after exchange with acetone, by gas chromatography-mass spectrometry. The combination of our 18O method and the 2H method of Yang et al. allows one to measure energy expenditure via "doubly labeled" water (2H(2)O + H(2)18O), using small samples of body fluids. These techniques were used to measure energy expenditure in mice, in which the 18O enrichment of body water can be monitored down to 0.025%.
Department of Nutrition, Case Western Reserve University, School of Medicine, Cleveland, Ohio 44106-4906, USA.
Daniel Z
DZ
Brunengraber
Brendan J
BJ
McCabe
Jill
J
Katanik
Stephen F
SF
Previs
D000818:Animals; D004734:Energy Metabolism; D008401:Gas Chromatography-Mass Spectrometry; D008297:Male; D051379:Mice; D010755:Organophosphates / Q000378:metabolism*; D010103:Oxygen Isotopes / Q000097:blood / Q000378:metabolism* / Q000652:urine; D014867:Water / Q000378:metabolism*
D010755:Organophosphates; D010103:Oxygen Isotopes; D014867:Water; C003715:trimethyl phosphate