A family of nanostructured polyaniline (PANI) materials including polystyrene (PS)/PANI core/shell particles, PANI hollow spheres, PANI/PS nanocomposite and nanoporous PANI, were conveniently prepared by surface charge control of PS nanoparticle templates which resulted in different polymer growth mechanisms when PANI was electropolymerized around the templates.

A biosensor with improved performance was developed through the immobilization of horseradish peroxidase (HRP) onto electropolymerized polyaniline (PANI) films doped with carbon nanotubes (CNTs). The effects of electropolymerization cycle and CNT concentration on the response of the biosensor toward H2O2 were investigated. It was found that the application of CNTs in the biosensor system could increase the amount and stability of the immobilized enzyme, and greatly enhanced the biosensor response. Compared with the biosensor without CNTs, the proposed biosensor exhibited enhanced stability and approximately eight-fold sensitivity. A linear range from 0.2 to 19 μM for the detection of H2O2 was observed for the proposed biosensor, with a detection limit of 68 nM at a signal-to-noise ratio of 3 and a response time of less than 5 s.

The characterization of a polyclonal antibody produced via immunization with an [Os(bpy)2dcbpy] hapten is described. Bpy is 2,2′-bipyridine and dcbpy is 2,2′-bipyridine-4,4′-dicarboxylic acid. The cross-reactivity of the antibody for the Ru(II) analogue of the hapten was also investigated. Large increases in the emission and luminescent lifetime of a series of Os and Ru complexes were observed on binding of the antibody. Association equilibrium constants were derived from luminescence titration data and were found to be 5.6 × 108 and 5.0 × 108 M−1 for [Os(bpy)2dcbpy] and [Ru(bpy)2dcbpy], respectively. Spectroscopic changes were likely due to the exclusion of H2O from the complex/antibody binding cleft and blocking of vibrational relaxation pathways of the Os/Ru excited state. D2O/H2O experiments confirmed that the antibody protected approx. 82% of [Os(bpy)2dcbpy] and 80% of [Ru(bpy)2dcbpy] from excited state deactivation by the aqueous solvent.

With lower limits of detection and increased stability constantly being demanded of biosensor devices, characterisation of the constituent layers that make up the sensor has become unavoidable, since this is inextricably linked with its performance. This work describe the optimisation and characterisation of two aspects of sensor performance: a conductive polymer layer (polyaniline) and the immobilised protein layer. The influence of the thickness of polyaniline films deposited electrochemically onto screen-printed electrode surfaces is described in this work in terms of its influence on a variety of amperometric sensor performance characteristics: time to reach steady state, charging current, catalytic current, background current and signal/background ratios. The influence of polymer film thickness on the conductivity and morphology of finished films is also presented.An electrostatic method of protein immobilisation is used in this work and scanning electron microscopy in conjunction with gold-labelled antibodies and back-scattered electron detection has enabled the direct visualisation of individual groups of proteins on the sensor surface. Such information can provide an insight into the performance of sensors under influence of increasing protein concentrations.

This work investigates polyaniline (PANI) nanoparticles, (synthesised using dodecylbenzenesulphonic acid (DBSA) as a dopant), as a novel, highly processable, non-diffusional mediating species in an enzyme biosensing application. These nanoparticles are readily dispersed in aqueous media which helps overcome some of the processability issues traditionally associated with polyaniline. Modification of screen-printed electrodes was readily achieved these aqueous nanoparticle dispersions, where the nanoparticles were simply cast by a drop-coating method onto the surface. After suitable pH adjustment, it was shown that horseradish peroxidase (HRP) enzyme could be added to the dispersion, and cast simultaneously with the conducting polyaniline. This effective fabrication method involves no electrochemical steps, and as such is easily amenable to mass production. The feasibility of casting enzyme with polyaniline nanoparticles is demonstrated in this short communication. More accurate deposition of protein-containing inks onto screen-printed carbon working electrodes could in the future transfer the drop-coating protocol from manual deposition to large-scale production by mechanical methods such as ink-jet printing.

8-Oxo-7,8-dihydroguanine (8-oxoGua), an important biomarker of DNA damage in oxidatively generated stress, is highly reactive towards further oxidation. Much work has been carried out to investigate the oxidation products of 8-oxoGua by one-electron oxidants, singlet oxygen, and peroxynitrite. This report details for the first time, the iron- and copper-mediated Fenton oxidation of 8-oxoGua and 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodGuo). Oxidised guanidinohydantoin (Ghox) was detected as the major product of oxidation of 8-oxoGua with iron or copper and hydrogen peroxide, both at pH 7 and pH 11. Oxaluric acid was identified as a final product of 8-oxoGua oxidation. 8-oxodGuo was subjected to oxidation under the same conditions as 8-oxoGua. However, dGhox was not generated. Instead, spiroiminodihydantoin (Sp) was detected as the major product for both iron and copper mediated oxidation at pH 7. It was proposed that the oxidation of 8-oxoGua was initiated by its one-electron oxidation by the metal species, which leads to the reactive intermediate 8-oxoGua˙+, which readily undergoes further oxidation. The product of 8-oxoGua and 8-oxodGuo oxidation was determined by the 2′-deoxyribose moiety of the 8-oxodGuo, not whether copper or iron was the metal involved in the oxidation.

This study presents the use of complementary colorimetric and amperometric techniques to measure the quantity of protein or enzyme immobilised onto a carbon paste electrode modified with a layer of electrodeposited polyaniline. By applying a solution of bovine serum albumin at 0.75 mg/ml, efficient blocking of the electrode from electroactive species in the bulk solution could be achieved. When the horseradish peroxidase was immobilised on the electrode, optimal amperometric responses from hydrogen peroxide reduction were achieved at approximately the same concentration. The mass of enzyme immobilised at this solution concentration was determined by a colorimetric enzyme assay to be equivalent to the formation of a protein monolayer. Under these conditions, amperometric responses from the immobilised layer are maximised and non-specific bulk solution interactions are minimised. At higher immobilised protein concentrations, diminished amperometric responses may be due to inhibited diffusion of hydrogen peroxide to enzyme which is in electronic communication with the electrode surface, or impeded electron transfer.

Cytochrome P450cam (CYP101) bioelectrodes suitable for application in organic phases were prepared from genetically engineered CYP101 and vesicular dispersions of didodecyldimethylammonium bromide. The amperometric biosensor system was characterised under anaerobic conditions by cyclic and square-wave voltammetric methods. Cyclic- and square-wave-voltammetry studies showed that the biosensors exhibited direct reversible electron transfer between the haem iron atom and the glassy carbon electrode surface. The formal redox potential estimated for the electrode in acetonitrile was −380 mV/Ag–AgCl. The formal potential shifted anodically as the organic phase biosensor responded irreversibly to substrate (camphor) under anaerobic and aerobic conditions in acetonitrile. Differential pulse analysis of the reactivities of the CYP101 enzyme electrode confirmed the square-wave voltammetry result, which showed that the binding of substrate decreased the redox potential necessary for initiating the monooxygenation reaction of cytochrome P450cam.

A multi-dimensional column chromatographic method employing UV spectrometric detection was optimised and successfully used in a comparative bio-availability study of aspirin obtained from different commercially available oral dosage forms. Sample clean-up was achieved by on-line solid-phase extraction. In this study, the bioavailability of aspirin was compared in plain aspirin tablets, chewed tablets, effervescent tablets and Enteric-coated aspirin tablets. Blood samples were taken at frequent intervals after single dosing in ten healthy volunteers, the plasma samples were first treated with physostigmine sulphate to minimise enzymatic hydrolysis of aspirin to salicylate. The results showed the measured Tmax, Cmax and AUC was significantly higher for soluble aspirin than for the other formulations and the t1/2 was shorter. This indicates the rapid absorption of aspirin from a soluble formulation compared with that from the other formulations. These differences suggest that the soluble formulation could be the aspirin of choice to treat patients suspected to be at high risk of myocardial infarction. The method performs, in a single step, an efficient extraction and clean-up of aspirin from human plasma. The calibration graph was linear over the calibration range 0.2–12 μg ml−1 plasma with a limit of detection of 0.1 μg ml−1. The intra- and inter-assay coefficients of variation were less than 6% and the recoveries ranged from 86 to 98%. The proposed method combines the advantages of being simple and selective in the presence of other potential interfering drugs and is suitable for routine analyses to obtain valuable information about the clinical effects of the drug and its use in prevention treatments of acute myocardial infarction. The whole procedure takes ∼7 min and is in agreement with other conventional methods.

A family of nanostructured polyaniline (PANI) materials including polystyrene (PS)/PANI core/shell particles, PANI hollow spheres, PANI/PS nanocomposite and nanoporous PANI, were conveniently prepared by surface charge control of PS nanoparticle templates which resulted in different polymer growth mechanisms when PANI was electropolymerized around the templates.