13C isotopomer analysis of glutamate by J-resolved heteronuclear single quantum coherence spectroscopy.
13C NMR isotopomer analysis is a powerful method for measuring metabolic fluxes through pathways intersecting in the tricarboxylic acid cycle. However, the inherent insensitivity of 13C NMR spectroscopy makes application of isotopomer analysis to small tissue samples (mouse tissue, human biopsies, or cells grown in tissue culture) problematic. (1)H NMR is intrinsically more sensitive than 13C NMR and can potentially supply the same information via indirect detection of 13C providing that isotopomer information can be preserved. We report here the use of J-resolved HSQC (J-HSQC) for 13C isotopomer analysis of tissue samples. We show that J-HSQC reports isotopomer multiplet patterns identical to those reported by direct 13C detection but with improved sensitivity.
Burgess
S C
SC
Department of Chemistry, University of Texas at Dallas, TX 75235, USA.
Carvalho
R A
RA
Merritt
M E
ME
Jones
J G
JG
Malloy
C R
CR
Sherry
A D
AD
D000818:Animals; D002244:Carbon / Q000737:chemistry*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D018698:Glutamic Acid / Q000032:analysis; D006859:Hydrogen / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D051379:Mice; D015425:Myocardial Reperfusion; D010477:Perfusion; D012680:Sensitivity and Specificity; D013997:Time Factors
D018698:Glutamic Acid; D002244:Carbon; D006859:Hydrogen