Optimization of hydroxyl radical formation using TiO2 as photocatalyst by response surface methodology

The determination of the optimum parameters for hydroxyl radicals (OH) formation by a TiO₂ solution has been investigated by measuring the emitted fluorescence after the reaction with terephthalic acid has occurred. After UV irradiation, the terephthalic acid was transformed into 2-hydroxyterephthalic acid whose fluorescence is directly proportional to the generated OH. Optimization of hydroxyl radicals’ formation using TiO₂ as catalyst was carried out by studying the effects of irradiation time, TiO₂ concentration and terephthalic acid concentration on the production of the fluorescent HTA with an experimental design. The aim of our research was to apply response surface methodology as a chemometric method for the optimization of the reaction conditions. The combination of irradiation time, TiO₂ concentration and terephthalic acid concentration was varied at designed points of a central composite rotatable design. The three factors were found to have a significant effect upon the reaction. The optimum conditions for the reaction achievement were estimated to be 10 min for the irradiation time, 25 μg mL⁻¹ TiO₂ concentration and 0.1 mmol L⁻¹ terephthalic acid concentration. Afterwards, using these parameters the method was applied for the determination of the ability of several plant extract samples to scavenge the formed OH.     

Development of a portable biosensor for screening neurotoxic agents in water samples

A high sensitive portable biosensor system capable of determining the presence of neurotoxic agents in water has been developed. The system consists of (i) a screen-printed electrode with acetylcholinesterase (AChE) immobilized on it, (ii) a self-developed portable potentiostat with an analog to digital (A/D) converter and a serial interface for transferring data to a portable PC and (iii) an own designed software, developed with Lab-Windows CVI, used to record and process the measurements. The system has been developed to perform high precision amperometrical measurements with low drifts, low noise and a good reproducibility. In the configuration depicted, the percentage of AChE inhibition is proportional to the content of neurotoxic agents in a sample. This type of measurement is performed by the steady-state method from the first steady current (by a phosphate buffer solution) and the second steady current (by an enzymatic reaction produced by the addition of acetylthiocholine chloride to the solution). Validation was performed by analyzing spiked water samples containing pesticides. The design is specially suited for screening purposes, does not need sample preconcentration, is totally autonomous and suitable for the field detection of neurotoxic agents in water.     

Pluronics‐Stabilized Gold Nanoparticles: Investigation of the Structure of the Polymer–Particle Hybrid

Hybrid gold–polymer nanoparticles are obtained by self‐assembly of amphiphilic copolymers (Pluronics) in solutions containing preformed gold nanoparticles (diameter ca. 12 nm). Dynamic light scattering, TEM, cryo‐TEM, and small‐angle neutron scattering experiments with contrast variation are used to characterize the structure of the gold–polymer particles. Five Pluronics (F127, F68, F88, F108, P84) with different molecular weights and hydrophilic/hydrophobic balances are investigated. Gold nanoparticles are individually embedded within globules of polymer, even under conditions for which Pluronics micelles do not form in solution. The hybrid particles are several tens of nanometers in size (larger than micelles of the corresponding Pluronics), and the size can be tuned by changing the temperature.     

Hyperbranched polyamidoamine as stabilizer for catalytically active nanoparticles in water

Poly(amidoamine) hyperbranched polymers (HYPAM) were successfully used to synthesize platinum nanoparticles of ca. 1.8 nm and to stabilize these particles in water. Furthermore these hyperbranched-stabilized platinum nanoparticles proved to be effective and robust for hydrogenation reaction in water.     

Probing the Lipid Annular Belt by Gas‐Phase Dissociation of Membrane Proteins in Nanodiscs

Interactions between membrane proteins and lipids are often crucial for structure and function yet difficult to define because of their dynamic and heterogeneous nature. Here, we use mass spectrometry to demonstrate that membrane protein oligomers ejected from nanodiscs in the gas phase retain large numbers of lipid interactions. The complex mass spectra that result from gas‐phase dissociation were assigned using a Bayesian deconvolution algorithm together with mass defect analysis, allowing us to count individual lipid molecules bound to membrane proteins. Comparison of the lipid distributions measured by mass spectrometry with molecular dynamics simulations reveals that the distributions correspond to distinct lipid shells that vary according to the type of protein–lipid interactions. Our results demonstrate that nanodiscs offer the potential for native mass spectrometry to probe interactions between membrane proteins and the wider lipid environment.     

Aptamer-based zearalenone assay based on the use of a fluorescein label and a functional graphene oxide as a quencher

Microchimica Acta - A versatile and cost-effective aptamer-based fluorescence quenching assay is described for the detection of the mycotoxin zearalenone (ZEN). Exfoliated functional graphene oxide...     

Determination of Antioxidant Capacity by Using Xanthine Oxidase Bioreactor Coupled with Flow‐through H2O2 Amperometric Biosensor

A new flow system for antioxidant capacity (AOC) estimation, consisting of a bioreactor, containing immobilized xanthine oxidase (XOD), coupled with a H₂O₂ amperometric biosensor, based on Os‐wired horseradish peroxidase, was developed. The H₂O₂, resulting from the enzymatic reaction between xanthine (XA) and XOD, was amperometrically monitored at ?0.1 V vs. Ag/AgCl/KCl_sat, in order to avoid the electrochemical interferences. Two protocols were used to perform the AOC evaluation: “steady‐state”, when the antioxidant (AOX) was injected in the XA flow, and “transient state”, when XA and AOX were simultaneously injected in the carrier flow. The AOC of some commercial beverages were evaluated and compared with those obtained with 2,2‐diphenyl‐1‐picrylhydrazyl radical and Folin–Ciocalteu methods.     

Structural and functional characterisation of a biohybrid material based on acetylcholinesterase and layered double hydroxides

This work describes the use of layered double hydroxides (LDHs) for the immobilisation of acetylcholinesterase (AChE) on insulator/semiconductor solid supports. Different LDHs have been synthesised by a co-precipitation method. Afterwards, biohybrid materials based on AChE-LDH mixtures have been produced using wild and recombinant enzymes. Spectroscopic techniques have confirmed the LDH phase identity and the links created between the LDH and AChE. Spectrophotometric assays have demonstrated that most of the biohybrid materials are functional and stable. Several configurations have been used for AChE immobilisation. The highest catalytic responses have been observed when using wild enzyme and immobilising AChE-LDH mixtures on LDHs previously deposited on the solid supports. LDHs have been demonstrated to be suitable host matrices for AChE immobilisation on electrodes for the subsequent development of electrochemical biosensors.     

Novel Amperometric Hydrogen Peroxide Biosensor Based on Horseradish Peroxidase Azide Covalently Immobilized on Ethynyl‐Modified Screen‐Printed Carbon Electrode via Click Chemistry

A novel, simple and versatile protocol for covalent immobilization of horseradish peroxidase (HRP) on screen‐printed carbon electrode (SPCE) based on the combination of diazonium salt electrografting and click chemistry has been successfully developed. The ethynyl‐terminated monolayers are obtained by diazonium salt electrografting, then, in the presence of copper (I) catalyst, the ethynyl modified surfaces reacted efficiently and rapidly with horseradish peroxidase bearing an azide function (azido‐HRP), thus forming a covalent 1,2,3‐triazole linkage by means of click chemistry. All the experimental results suggested that HRP was immobilized onto the electrode surface successfully without denaturation. Furthermore, the immobilized HRP showed a fast electrocatalytic reduction for H₂O₂. A linear range from 5.0 to 50.0 µM in a phosphate buffer (pH 5.5) with detection limit of 0.50 µM and sensitivity of 0.23 nA/µM were obtained. The heterogeneous electron transfer rate constant K_ct was 1.52±0.22 s^?1 and the apparent Michaelis? Menten constant was calculated to be 0.028 mM. The HRP‐functionalized electrode demonstrated a good reproducibility and long‐term stability.