Electrochemical sensor for catechol and dopamine based on a catalytic molecularly imprinted polymer-conducting polymer hybrid recognition element.

One of the difficulties with using molecularly imprinted polymers (MIPs) and other electrically insulating materials as the recognition element in electrochemical sensors is the lack of a direct path for the conduction of electrons from the active sites to the electrode. We have sought to address this problem through the preparation and characterization of novel hybrid materials combining a catalytic MIP, capable of oxidizing the template, catechol, with an electrically conducting polymer. In this way a network of "molecular wires" assists in the conduction of electrons from the active sites within the MIP to the electrode surface. This was made possible by the design of a new monomer that combines orthogonal polymerizable functionality; comprising an aniline group and a methacrylamide. Conducting films were prepared on the surface of electrodes (Au on glass) by electropolymerization of the aniline moiety. A layer of MIP was photochemically grafted over the polyaniline, via N,N'-diethyldithiocarbamic acid benzyl ester (iniferter) activation of the methacrylamide groups. Detection of catechol by the hybrid-MIP sensor was found to be specific, and catechol oxidation was detected by cyclic voltammetry at the optimized operating conditions: potential range -0.6 V to +0.8 V (vs Ag/AgCl), scan rate 50 mV/s, PBS pH 7.4. The calibration curve for catechol was found to be linear to 144 microM, with a limit of detection of 228 nM. Catechol and dopamine were detected by the sensor, whereas analogues and potentially interfering compounds, including phenol, resorcinol, hydroquinone, serotonin, and ascorbic acid, had minimal effect (< or = 3%) on the detection of either analyte. Non-imprinted hybrid electrodes and bare gold electrodes failed to give any response to catechol at concentrations below 0.5 mM. Finally, the catalytic properties of the sensor were characterized by chronoamperometry and were found to be consistent with Michaelis-Menten kinetics.

Chimeric polymers formed from a monomer capable of free radical, oxidative and electrochemical polymerisation.

A new monomer, which incorporates both aniline and methacrylamide functional groups, was shown to possess orthogonal polymerisation behaviour to produce conjugated polyaniline suitable for a wide range of applications.

Imprinted polymer-modified hanging mercury drop electrode for differential pulse cathodic stripping voltammetric analysis of creatine.

The molecularly imprinted polymer [poly(p-aminobenzoicacid-co-1,2-dichloroethane)] film casting was made on the surface of a hanging mercury drop electrode by drop-coating method for the selective and sensitive evaluation of creatine in water, blood serum and pharmaceutical samples. The molecular recognition of creatine by the imprinted polymer was found to be specific via non-covalent (electrostatic) imprinting. The creatine binding could easily be detected by differential pulse, cathodic stripping voltammetric signal at optimised operational conditions: accumulation potential -0.01 V (versus Ag/AgCl), polymer deposition time 15s, template accumulation time 60s, pH 7.1 (supporting electrolyte< or =5 x 10(-4)M NaOH), scan rate 10 mV s(-1), pulse amplitude 25 mV. The modified sensor in the present study was found to be highly reproducible and selective with detection limit 0.11 ng mL(-1) of creatine. Cross-reactivity studies revealed no response to the addition of urea, creatinine and phenylalanine; however, some insignificant magnitude of current was observed for tryptophan and histidine in the test samples.

Molecularly imprinted polymer-based solid-phase extraction combined with molecularly imprinted polymer-based sensor for detection of uric acid.

The development of an easy-to-use, rapid, robust and inexpensive technique is required which can measure the basal concentration of uric acid (UA) lower than 1.0 x 10(-7)M ( approximately 0.017 mgL(-1)) in biological samples to attend the problem of hypouricemia. In the present work an artificial receptor for UA, silica gel-bonded molecularly imprinted polymer (MIP), was used as a sorbent for molecularly imprinted solid-phase extraction (MISPE) in column chromatography. The use of a sensor based on a MIP-modified hanging mercury drop electrode (HMDE), as reported from our laboratory, could estimate UA with detection limit as low as 0.024 mgL(-1) under the optimized conditions of differential pulse, cathodic stripping voltammetric (DPCSV) measurement. However, in the current investigation, with the use of the combination of MISPE followed by detection with a MIP-based HMDE sensor, the minimum detectable concentration could go down to 0.0008 mgL(-1) (RSD=0.63%, S/N=3). The same MIP receptor for both MISPE and the corresponding sensor was able to enhance the preconcentration of analyte substantially so as to attain the desired level of sensitivity; and that to without any interference (cross-reactivity) from other structurally related analogues including the major interferent like ascorbic acid prevalent in the aqueous environment of biological samples.

Microplates with adaptive surfaces.

Here we present a new and versatile method for the modification of the well surfaces of polystyrene microtiter plates (microplates) with poly(N-phenylethylene diamine methacrylamide), (poly-NPEDMA). The chemical grafting of poly-NPEDMA to the surface of microplates resulted in the formation of thin layers of a polyaniline derivative bearing pendant methacrylamide double bonds. These were used as the attachment point for various functional polymers through photochemical grafting of various, for example, acrylate and methacrylate, polymers with different functionalities. In a model experiment, we have modified poly-NPEDMA-coated microplates with a small library of polymers containing different functional groups using a two-step approach. In the first step, double bonds were activated by UV irradiation in the presence of N,N-diethyldithiocarbamic acid benzyl ester (iniferter). This enabled grafting of the polymer library in the second step by UV irradiation of solutions of the corresponding monomers in the microplate wells. The uniformity of coatings was confirmed spectrophotometrically, by microscopic imaging and by contact angle measurements (CA). The feasibility of the current technology has been shown by the generation of a small library of polymers grafted to the microplate well surfaces and screening of their affinity to small molecules, such as atrazine, a trio of organic dyes, and a model protein, bovine serum albumin (BSA). The stability of the polymers, reproducibility of measurement, ease of preparation, and cost-effectiveness make this approach suitable for applications in high-throughput screening in the area of materials research.

Molecularly imprinted solid-phase extraction combined with molecularly imprinted polymer-sensor: a diagnostic tool applicable to creatine deficiency syndrome.

Primary creatine deficiency syndromes (CDS) are a new group of disorders caused by guanidinoacetate methyltransferase (GAMT) deficiency, which affects endogenous creatine biosynthesis with depletion of body creatine. A deficiency in creatine can be corrected by treatment with oral creatine supplementation and this necessitates a simple and sensitive screening method for early detection of creatine in dilute physiologic fluids. In this work an artificial receptor, molecularly imprinted polymer (MIP), for creatine was used both as a material for solid-phase extraction (SPE) and as a sensing element in a voltammetric sensor. Using the combination of molecularly imprinted solid-phase extraction (MISPE) with a complementary MIP sensor, the minimum detectable amount was found to be 0.0015 ng mL(-1) (RSD = 1.3%, S/N = 3). The MISPE-MIP sensor combination provided up to 60-fold preconcentration, which was more than sufficient for achieving the required quantification limit 50 ng mL(-1) (or 0.0025 ng mL(-1) after 2 x 10(4)-fold dilution) for creatine in human blood serum.