Covalent cucurbit[7]uril-dye conjugates for sensing in aqueous saline media and biofluids.

Non-covalent chemosensing ensembles of cucurbit[n]urils (CBn) have been widely used in proof-of-concept sensing applications, but they are prone to disintegrate in saline media, e.g. biological fluids. We show here that covalent cucurbit[7]uril-indicator dye conjugates are buffer- (10× PBS buffer) and saline-stable (up to 1.4 M NaCl) and allow for selective sensing of Parkinson's drug amantadine in human urine and saliva, where the analogous non-covalent CB7⊃dye complex is dysfunctional. The in-depth analysis of the covalent host-dye conjugates in the gas-phase, and deionized versus saline aqueous media revealed interesting structural, thermodynamic and kinetic effects that are of general interest for the design of CBn-based supramolecular chemosensors and systems. This work also introduces a novel high-affinity indicator dye for CB7 through which fundamental limitations of indicator displacement assays (IDA) were exposed, namely an impractical slow equilibration time. Unlike non-covalent CBn⊃dye reporter pairs, the conjugate chemosensors can also operate through a SN2-type guest-dye exchange mechanism, which shortens assay times and opens new avenues for tailoring analyte-selectivity.

Electrochemical Examination of the Structure of Thin Hydrogel Layers Anchored to Regular and Microelectrode Surfaces.

For the examination of hydrogel structure, thin layers of thermoresponsive gels based on poly(N-isopropylacrylamide) (pNIPA) and copolymer poly(N-isopropylacrylamide-co-sodium acrylate) (p(NIPA-co-AS)) were successfully anchored to microelectrode and regular electrode surfaces using the electrochemically induced free radical polymerization. The obtained layers were stable and covered the entire surface of the electrodes. Electroactive probes 1,1'-ferrocenedimethanol (Fc(CH2OH)2) and synthesized derivatives of ferrocene modified with polyethylene glycol units (Fc-PEGn) of various length (n = 4, 9, 75, and 135) were employed for studying the volume phase transition of the thin hydrogel layers and for the determination of their structural parameters. The quantitative information on the structural parameters of the hydrogel layers was derived from the obstruction model for diffusion using the voltammetrically determined diffusion coefficients for the model redox probe Fc(CH2OH)2. An approach to the determination of the effective radii of the gel openings (channels) for pNIPA and p(NIPA-co-AS) microlayers was developed. The obtained results were matched with the experimental results and allowed derivation of quantitative conclusions. The voltammograms obtained with modified electrodes in solutions containing Fc-PEG4, Fc-PEG9, and Fc-PEG75 were well defined and of appropriate height. However, the voltammograms recorded for Fc-PEG135, the hydrodynamic radius of which exceeded the size of the gel channels, were at the baseline level.