A potentiometric method to determine ionophore complex formation constants in solvent polymeric membrane phases, proposed originally by Russian researchers, is critically evaluated and compared to other established methods. It requires membrane potential measurements on two-layer sandwich membranes, where only one side contains the ionophore. The resulting initial membrane potential reflects the ion activity ratio at both aqueous phase--membrane interfaces and can be conveniently used to calculate complex formation constants in situ. This method is potentially useful, since it does not require the use of a reference ion or second ionophore in the measurement. In this paper, the five ionophores valinomycin, BME-44, ETH 2120, tert-butylcalix[4]arene tetraethyl ester, and S,S'-methylenebis(diisobutyldithiocarbamate) are characterized in poly(vinyl chloride) (PVC) plasticized with dioctyl sebacate (DOS) and compared with other established methods. The resulting formation constants correspond well to literature values. The influence of varying membrane concentrations and different anionic site additives is studied and found to be relatively small. Experiments are also performed with and without lipophilic inert electrolytes and with ionophore-free sandwich membranes to illustrate the effect of ion pairing and the membrane internal diffusion potential on the response of such sandwich membranes. These experiments suggest that ions are completely associated in PVC-DOS membranes, but that such ion pairs are rather nonspecific. Diffusion potentials seem to play a minor role with these systems. The results are explained with theory. This work indicates that the characterization of electrically charged ionophores, anion-selective ionophores, and ionophores in membrane matrixes other than PVC plasticized with DOS may now be experimentally accessible.
Molecular beacons are a new class of fluorescent probes that can report the presence of specific nucleic acids with high sensitivity and excellent specificity. In addition to their current wide applications in monitoring the progress of polymerase chain reactions, their unique properties make them promising probes for the detection and visualization of target biomolecules in living cells. This article is focused on our recent research in exploring the potential of using molecular beacon for living-cell studies in three important areas: the monitoring of mRNA in living cells, the development of ultrasmall DNA/RNA biosensors, and the novel approach of combining molecular beacon's signal transduction mechanism with aptamer's specificity for real-time protein detection. These applications demonstrate molecular beacon's unique properties in bioanalysis and bioassay development.
Lipophilic inert electrolytes, i.e., salts without ion-exchange properties, may influence the selectivity of ionophore-based liquid membrane electrodes by affecting the activity coefficients in the organic phase. It is expected by a theoretical model that the addition of a lipophilic salt renders the ion-selective electrode more selective for divalent over monovalent ions. These predictions are confirmed with Ca(2+)-responsive membranes containing the ionophores ETH 2120, ETH 1001, and ETH 129. The effect is especially pronounced with nonpolar membrane phases containing a low concentration of charged species, where up to 2 orders of magnitude selectivity improvement is observed.
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