A sample preparation protocol for 1H nuclear magnetic resonance studies of water-soluble metabolites in blood and urine.
We describe a general protocol for preparing protein-containing biofluids for (1)H nuclear magnetic resonance (NMR) metabolomic studies. In this protocol, untreated samples are diluted in deuterated solvents to precipitate proteins and recover metabolites quantitated relative to standard reference compounds such as 3-trimethylsilylpropionic acid (TSP) and 2,2-dimethyl-2-silapentane-5-sulfonic acid (DSS). The efficacy of this protocol was tested using a bovine serum albumin/metabolite mix and human serum samples. This sample preparation method can be readily applied to any protein-containing biofluid for (1)H NMR studies.
Sheedy
John R
JR
Department of Medicine, Royal Melbourne and Western Hospitals, University of Melbourne, Footscray, Victoria 3011, Australia. jsheedy@unimelb.edu.au
Ebeling
Peter R
PR
Gooley
Paul R
PR
McConville
Malcolm J
MJ
D053000:Analytic Sample Preparation Methods / Q000379:methods*; D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods*; D002417:Cattle; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism*; D012995:Solubility; D016482:Urinalysis / Q000379:methods*; D014867:Water / Q000737:chemistry*
D011506:Proteins; D014867:Water