Separation of intra- and extracellular lactate NMR signals using a lanthanide shift reagent.
This work describes the use of [Pr-DO3A] as a shift reagent for differentiating intra- and extracellular L-lactate resonance in (1)H- and (13)C-NMR spectra. DO3A acts as heptadentate ligand towards lanthanide(III) ions, leaving two coordination vacancies for the coordination of the L-lactate ion. The exchange between free and [Pr-DO3A]-bound L-lactate is fast on the NMR timescale, thus yielding a paramagnetically shifted L-lactate signal for the substrate in the compartment containing the paramagnetic chelate. The evaluation of the method was carried out on a model system based on sealed ghosts from human red blood cells.
Aime
Silvio
S
Dipartimento di Chimica IFM, Università di Torino, Torino, Italy. aime@ch.unito.it
Botta
Mauro
M
Mainero
Valentina
V
Terreno
Enzo
E
D000085:Acetates; D004910:Erythrocyte Membrane; D006573:Heterocyclic Compounds, 1-Ring; D006801:Humans; D007202:Indicators and Reagents; D019344:Lactic Acid / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy
C436944:1,4,7-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane; D000085:Acetates; D006573:Heterocyclic Compounds, 1-Ring; D007202:Indicators and Reagents; D019344:Lactic Acid