Whole Blood Metabolomics by1H NMR Spectroscopy Provides a New Opportunity To Evaluate Coenzymes and Antioxidants.
Conventional human blood metabolomics employs serum or plasma and provides a wealth of metabolic information therein. However, this approach lacks the ability to measure and evaluate important metabolites such as coenzymes and antioxidants that are present at high concentrations in red blood cells. As an important alternative to serum/plasma metabolomics, we show here that a simple1H NMR experiment can simultaneously measure coenzymes and antioxidants in extracts of whole human blood, in addition to the nearly 70 metabolites that were shown to be quantitated in serum/plasma recently [ Anal. Chem. 2015 , 87 , 706 - 715 ]. Coenzymes of redox reactions: oxidized/reduced nicotinamide adenine dinucleotide (NAD+and NADH) and nicotinamide adenine dinucleotide phosphate (NADP+and NADPH); coenzymes of energy including adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP); and antioxidants, the sum of oxidized and reduced glutathione (GSSG and GSH) can be measured with essentially no additional effort. A new method was developed for detecting many of these unstable species without affecting other blood/blood plasma metabolites. The identities of coenzymes and antioxidants in blood NMR spectra were established combining 1D/2D NMR techniques, chemical shift databases, pH measurements and, finally, spiking with authentic compounds. This is the first study to report identification of major coenzymes and antioxidants and quantify them, simultaneously, with the large pool of other metabolites in human blood using NMR spectroscopy. Considering that the levels of coenzymes and antioxidants represent a sensitive measure of cellular functions in health and numerous diseases, the NMR method presented here potentially opens a new chapter in the metabolomics of blood.
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0000-0002-0544-7464
GA
Nagana Gowda
Fred Hutchinson Cancer Research Center , Seattle, Washington 98109, United States.
Daniel
D
Raftery
D000244:Adenosine Diphosphate / Q000097:blood; D000249:Adenosine Monophosphate / Q000097:blood; D000255:Adenosine Triphosphate / Q000097:blood; D000975:Antioxidants / Q000032:analysis* / Q000737:chemistry; D001224:Aspartic Acid / Q000097:blood; D003067:Coenzymes / Q000097:blood*; D062126:Databases, Chemical; D005978:Glutathione / Q000097:blood; D006801:Humans; D006863:Hydrogen-Ion Concentration; D055432:Metabolomics; D009243:NAD / Q000097:blood; D009249:NADP / Q000097:blood; D010084:Oxidation-Reduction; D066244:Proton Magnetic Resonance Spectroscopy
D000975:Antioxidants; D003067:Coenzymes; D009243:NAD; D001224:Aspartic Acid; D000249:Adenosine Monophosphate; D009249:NADP; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D005978:Glutathione