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August 1, 1997

Direct observation of resolved intracellular and extracellular water signals in intact human red blood cells using 1H MAS NMR spectroscopy

Key Points

This research aims to directly observe intracellular and extracellular water signals in intact human red blood cells using NMR spectroscopy.
High resolution 400 MHz 1H NMR spectroscopy with magic angle spinning was used.
Paramagnetic Mn-EDTA complex and variations in H2O/D2O ratios were employed to measure relaxation times.
Two distinct water resonances were identified and assigned to intracellular and extracellular sources.
Two water resonances were detected, separated by 15 Hz, corresponding to intracellular (17.5 Hz linewidth) and extracellular water (4.6 Hz linewidth).
This study provides the first direct observation of intracellular water using the 1H MAS NMR spectroscopy technique.
The approach allows for the investigation of substance compartmentation and kinetics in cells.

Abstract

High resolution 400 MHz 1H NMR spectra of red blood cell suspensions when measured using magic angle spinning (MAS) show two water resonances separated by 15 Hz. Based on addition of a paramagnetic Mn-EDTA complex, measurement of relaxation times and variation of extracellular H2O/D2O ratios, these have been assigned as intracellular (linewidth 17.5 Hz) and extracellular water (linewidth 4.6 Hz). This is the first direct observation of intracellular water using NMR spectroscopy and the 1H MAS NMR spectroscopic approach offers the possibility of studying directly the compartmentation of substances in cells and kinetics of molecular transport.

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