Protein Interference with Ion-Selective Electrode Measurement Depends on Reference Electrode Composition and Design

Abstract

There is controversy about whether protein interferes with ion measurements using ion-selective electrodes. We have investigated the effects of changes in the salt-bridge composition of five commercially available analysers with open, membrane-restricted or porous frit-restricted reference electrode junctions on measurements of an albumin solution prepared by gel filtration. When the manufacturers' salt bridges were used, instruments with open or membrane-restricted junctions showed apparent increases in the activity of ionized calcium, sodium and potassium in the presence of protein. When the hypertonic bridge solutions were replaced with 150 mmol/L potassium chloride this increase disappeared. The instrument with a porous frit-restricted junction showed no protein effect, but its response to changes in sample sodium chloride concentration in protein-free solution suggested that its junction was functionally equivalent to that formed with an isotonic sodium chloride bridge. Our results emphasize that liquid junction design and composition affect ion measurements in protein-containing solutions and suggest that the use of hypertonic bridge solutions for biological samples needs to be re-examined.