Intra- and extraerythrocyte pH at 37°C and during long term storage at 4°C

31P NMR measurements and an electrochemical model of the system

J. E. Raftos*, B. E. Chapman, P. W. Kuchel, V. A. Lovric, I. M. Stewart

*Corresponding author for this work

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Abstract

A 31P NMR method based on pH dependent variation of the chemical-shift-difference between the resonances of orthophosphate and methylphosphonate was used to measure simultaneously intracellular pH (pHi) and extracellular pH (pHc) during long term storage of erythrocytes; pH was determined at both 4°C and 37°C. An equation describing the equilibrium distribution of membrane-permeant ions was derived by consideration of the electrochemical and osmotic constraints in the RBC suspension. Calculations using the model-equation and the measured pHi yielded the Donnan ratio and therefore pHo; the relationship between experimentally determined pHi and pHo values was accurately predicted by the model. Sensitivity analysis of the model-equation revealed that the observed increase in transmembrane pH gradient during storage is principally due to the alteration of the total net charge of intracellular (poly)-anions.

Original languageEnglish
Pages (from-to)251-268
Number of pages18
JournalHaematologia
Volume19
Issue number4
Publication statusPublished - 1986

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    Raftos, J. E., Chapman, B. E., Kuchel, P. W., Lovric, V. A., & Stewart, I. M. (1986). Intra- and extraerythrocyte pH at 37°C and during long term storage at 4°C: 31P NMR measurements and an electrochemical model of the system. Haematologia, 19(4), 251-268.