Abstract
A method is described for determining the intracellular pH of intact erythrocytes by 1H NMR. The determination is based on the pH dependence of the chemical shifts of resonances for carbon-bonded protons of an indicator molecule (imidazole) in intact cells. The imidazole is introduced into the erythrocytes by incubation in an isotonic saline solution of the indicator. The pH dependence of the chemical shifts of the imidazole resonances is calibrated from 1H NMR spectra of the imidazole-containing red cell lysates whose pH is varied by the addition of acid or base and measured directly with a pH electrode. To reduce in intensity or eliminate the much more intense envelope of resonances from the hemoglobin, the 1H NMR measurements are made by either the spin-echo Fourier transform technique or by the transfer-of-saturation by cross-relaxation method.
| Original language | English |
|---|---|
| Pages (from-to) | 423-432 |
| Number of pages | 10 |
| Journal | Analytical Biochemistry |
| Volume | 121 |
| Issue number | 2 |
| DOIs | |
| State | Published - Apr 1982 |
| Externally published | Yes |
Bibliographical note
Funding Information:We express our appreciation to the Hematology Division of the Department of Laboratory Medicine, University of Alberta Hospital, for providing the blood samples. This research was supported by an operating grant to D.L.R. from the Natural Sciences and Engineering Research Council and a Postdoctoral Fellowship to A.A.I. from the Alberta Heritage Foundation for Medical Research.
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology