Highly electroactive Co-ZnO/GO nanocomposite: Electrochemical sensing platform for oxytetracycline determination.

Antimicrobial residues in animal-derived foods have become a major source of concern around the world. Oxytetracycline (OTC), one of these antibiotics that belongs to the tetracycline family should be detected in these matrices. Nanostructured metal oxides have attracted a lot of scientific attention due to their special characteristics that can be exploited for creating innovative nanodevices. Therefore, in the present study, we report the fabrication of cobalt-doped ZnO/GO nanocomposites for OTC sensors using a simple and environmentally friendly method that does not require toxic solvents. Contact angle measurements, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and UV-Vis were used to confirm the successful fabrication of the Co-ZnO/GO nanocomposite and to determine the surface area, Structural, morphological features, chemical composition and purity of the nanocomposite. The electrochemical and electrocatalytic properties were recorded using cyclic voltammetry (CV), electrochemical impedance spectroscopy, and differential pulse voltammetry (DPV). Optimizing parameters such as scan rate, pH value, deposition time, and deposition potential, we achieve a wide linear concentration range from 10-12 M to 10-7 M, with an impressive detection limit of 1.6 10-13 M.Notably, our sensor exhibits remarkable selectivity, demonstrating its usefulness for the detection of oxytetracycline traces in real milk samples. These results emphasize the novelty and practical significance of our work and provide a promising avenue for the development of sensitive and selective electrochemical sensing platforms in various fields.

Mechanisms of Influenza Virus HA2 Peptide Interaction with Liposomes Studied by Dual-Wavelength MP-SPR.

A phospholipid-based liposome layer was used as an effective biomimetic membrane model to study the binding of the pH-dependent fusogenic peptide (E4-GGYC) from the influenza virus hemagglutinin HA2 subunit. To this end, a multiparameter surface plasmon resonance approach (MP-SPR) was used for monitoring peptide-liposome interactions at two pH values (4.5 and 8) by means of recording sensorgrams in real time without the need for labeling. Biotinylated liposomes were first immobilized as a monolayer onto the surface of an SPR gold chip coated with a streptavidin layer. Multiple sets of sensorgrams with different HA2 peptide concentrations were generated at both pHs. Dual-wavelength Fresnel layer modeling was applied to calculate the thickness (d) and the refractive index (n) of the liposome layer to monitor the change in its optical parameters upon interaction with the peptide. At acidic pH, the peptide, in its α helix form, entered the lipid bilayer of liposomes, inducing vesicle swelling and increasing membrane robustness. Conversely, a contraction of liposomes was observed at pH 8, associated with noninsertion of the peptide in the double layer of phospholipids. The equilibrium dissociation constant KD = 4.7 × 10-7 M of the peptide/liposome interaction at pH 4.5 was determined by fitting the "OneToOne" model to the experimental sensorgrams using Trace Drawer software. Our experimental approach showed that the HA2 peptide at a concentration up to 100 µM produced no disruption of liposomes at pH 4.5.

Exploring the benefits of surface analysis techniques to develop double multilayer transfer printing of J-Aggregates cyanine dyes by integrating L-b-L and µCp processes.

Layer-by-layer self-assembly (L-b-L assembly) makes possible to obtain polyelectrolyte multilayers (PEMs) and one of the polyelectrolytes could be replaced by a dye molecule to obtain multilayers which may exhibit optical properties of great interest. On the other hand, µCp has become a routine technique for the preparation of micro- and nanostructured surfaces. In our development in progress of a surface engineering strategy to transfer J-Agg cyanine dyes onto surfaces by integrating L-b-L process and µCp, this contribution highlights how surface analysis imaging techniques can bring valuable information for the development of the process involving a double Multilayers Transfer Printing (MTP) with a Moiré effect. Key parameters sustaining image interpretation are difference in deposit thickness (optical microscopy, atomic force microscopy, scanning electron microscopy), in roughness (atomic force microscopy and scanning electron microscopy), in charge effect (scanning electron microscopy) and the chemical contrast between unprinted and printed areas (time-of-flight secondary ion mass spectrometry).

Cleaner degreasing of sheepskins by the Yarrowia lipolytica LIP2 lipase as a chemical-free alternative in the leather industry.

Conventional degreasing of skins and hides in the leather industry requires high amounts of organic solvents and detergents that cause environmental issues. In this study, the LIP2 lipase from the yeast Yarrowia lipolytica (YLLIP2) was shown to be effective in degreasing sheepskins, thus reducing the amount of harmful chemicals. Using 6 mg of lipase/kg of raw skin, successful degreasing was achieved in only 15 min at pH 8 and 30°C. ToF-SIMS mass spectra of chemically and enzymatically treated sheepskins are consistent with a selective elimination process for the enzymatic treatment. Comparative SEM microscopy, ATR-FTIR spectroscopy and physicochemical analyses showed better properties of the enzymatically treated leather than those of the chemical treatment. Effluent physicochemical parameters showed that the enzymatic treatment is a cleaner degreasing operation. Altogether, this work opens new horizons to use the YLLIP2 lipase as a more efficient alternative in the leather industry.

Surface Plasmon Resonance Monitoring of Mono-Rhamnolipid Interaction with Phospholipid-Based Liposomes.

The interactions of mono-rhamnolipids (mono-RLs) with model membranes were investigated through a biomimetic approach using phospholipid-based liposomes immobilized on a gold substrate and also by the multiparametric surface plasmon resonance (MP-SPR) technique. Biotinylated liposomes were bound onto an SPR gold chip surface coated with a streptavidin layer. The resulting MP-SPR signal proved the efficient binding of the liposomes. The thickness of the liposome layer calculated by modeling the MP-SPR signal was about 80 nm, which matched the average diameter of the liposomes. The mono-RL binding to the film of the phospholipid liposomes was monitored by SPR and the morphological changes of the liposome layer were assessed by modeling the SPR signal. We demonstrated the capacity of the MP-SPR technique to characterize the different steps of the liposome architecture evolution, i.e., from a monolayer of phospholipid liposomes to a single phospholipid bilayer induced by the interaction with mono-RLs. Further washing treatment with Triton X-100 detergent left a monolayer of phospholipid on the surface. As a possible practical application, our method based on a biomimetic membrane coupled to an SPR measurement proved to be a robust and sensitive analytical tool for the detection of mono-RLs with a limit of detection of 2 µg mL-1.

Laccase-Based Biosensor Encapsulated in a Galactomannan-Chitosan Composite for the Evaluation of Phenolic Compounds.

Galactomannan, a neutral polysaccharide, was extracted from carob seeds and characterized. It was used for the first time for the fabrication of a laccase-based biosensor by the encapsulation of laccase in a chitosan+galactomannan composite. The fabricated biosensor was characterized by FTIR, scanning electron microscopy and cyclic voltammetry. The pyrocatechol detection was obtained by cyclic voltammetry measurements, through the detection of o-quinone at -0.447 V. The laccase activity was well preserved in the chitosan+galactomannan composite and the sensitivity of detection of pyrocatechol in the 10-16 M-10-4 M range was very high. The voltammetric response of the biosensor was stable for more than two weeks. To estimate the antioxidant capacity of olive oil samples, it was shown that the obtained laccase-based biosensor is a valuable alternative to the colorimetric Folin-Ciocalteu method.

Development of enteric polymer-based microspheres by spray-drying for colonic delivery of Lactobacillus rhamnosus GG.

Antibiotics are well-known disruptive elements of the intestinal microbiota and antibiotic-associated diarrhea appeared as the most common complication related with post-antibiotic dysbiosis. Lactobacillus rhamnosus GG (LGG) strain is very effective in preventing antibiotic-associated diarrhea in children and adults. However, as any probiotics, it is concerned by the loss of viability during storage and gastrointestinal transit. The aim of this study was to develop an encapsulation system suitable for the specific colonic delivery of LGG strain after oral administration. For this purpose, spray-dried Eudragit® S100 microparticles encapsulating LGG bacteria were developed by using an aqueous based spray-drying approach, avoiding the use of organic solvents. Carbohydrates were added to the formulation since they are widely used as protective agents of bacteria against the harmful effect of dehydration stress. Here, both Surface Enhanced Raman Scattering (SERS) and conventional plate count methods showed that carbohydrates increased the survival ratio of bacteria after spray-drying from 3 to more than 50%. Moreover, these protective agents ensured low residual moisture content thus providing great stability of the cells in the spray-dried powder during storage. Significant improvement of the cell viability in simulated gastro intestinal fluid (SGIF) was observed for encapsulated cells as compared with free LGG bacteria for which no viable cell was detectable after 1 h incubation in gastric fluid only. As a consequence, 4.5 × 107 CFU/g of encapsulated LGG were found viable after incubation of microparticles 1 h in Simulated Gastric Fluid followed by 6 h in Simulated Intestinal Fluid, corresponding to less than 3 log reduction of viable cells during the 7 h incubation in Simulated Gastro Intestinal Fluid. These results attested that the developed encapsulation system is suitable for its use as a bacteria carrier for specific colonic delivery.

Voltammetric Sensor Based on Molecularly Imprinted Chitosan-Carbon Nanotubes Decorated with Gold Nanoparticles Nanocomposite Deposited on Boron-Doped Diamond Electrodes for Catechol Detection.

Phenolic compounds such as catechol are present in a wide variety of foods and beverages; they are of great importance due to their antioxidant properties. This research presents the development of a sensitive and biocompatible molecular imprinted sensor for the electrochemical detection of catechol, based on natural biopolymer-electroactive nanocomposites. Gold nanoparticle (AuNP)-decorated multiwalled carbon nanotubes (MWCNT) have been encapsulated in a polymeric chitosan (CS) matrix. This chitosan nanocomposite has been used to develop a molecular imprinted polymers (MIP) in the presence of catechol on a boron-doped diamond (BDD) electrode. The structure of the decorated MWCNT has been studied by TEM, whereas the characterization of the sensor surface has been imaged by AFM, demonstrating the satisfactory adsorption of the film and the adequate coverage of the decorated carbon nanotubes on the electrode surface. The electrochemical response of the sensor has been analyzed by cyclic voltammetry (CV) where excellent reproducibility and repeatability to catechol detection in the range of 0 to 1 mM has been found, with a detection limit of 3.7 × 10-5 M. Finally, the developed sensor was used to detect catechol in a real wine sample.

Development of uncoated near-spherical gold nanoparticles for the label-free quantification of Lactobacillus rhamnosus GG by surface-enhanced Raman spectroscopy.

The Surface-enhanced Raman spectroscopy (SERS) method based on gold nanoparticles as SERS substrate was investigated for the label-free detection and quantification of probiotic bacteria that are widely used in various pharmaceutical formulations. Indeed, the development of a simple and fast SERS method dedicated to the quantification of bacteria should be very useful for the characterization of such formulations in a more convenient way than the usually performed tedious and time-consuming conventional counting method. For this purpose, uncoated near-spherical gold nanoparticles were developed at room temperature by acidic treatment of star-like gold nanoparticle precursors. In this study, we first investigated the influence of acidic treatment conditions on both the nanoparticle physicochemical properties and SERS efficiency using Rhodamine 6G (R6G) as "model" analyte. Results highlighted that an effective R6G Raman signal enhancement was obtained by promoting chemical effect through R6G-anion interactions and by obtaining a suitable aggregation state of the nanoparticles. Depending on the nanoparticle synthesis conditions, R6G SERS signals were up to 102-103-fold greater than those obtained with star-like gold nanoparticles. The synthesized spherical gold nanoparticles were then successfully applied for the detection and quantification of Lactobacillus rhamnosus GG (LGG). In that case, the signal enhancement was especially due to the combination of anion-induced chemical enhancement and nanoparticle aggregation on LGG cell wall consecutive to non-specific interactions. Both the simplicity and speed of the procedure, achieved under 30 min, including nanoparticle synthesis, sample preparation, and acquisition of SERS spectra, appeared as very relevant for the characterization of pharmaceutical formulations incorporating probiotics. Graphical abstract.

Highly Sensitive Voltammetric Glucose Biosensor Based on Glucose Oxidase Encapsulated in a Chitosan/Kappa-Carrageenan/Gold Nanoparticle Bionanocomposite.

In this work, an enzymatic sensor, based on a bionanocomposite film consisting of a polyelectrolyte complex (PEC) (Chitosan/kappa-carrageenan) doped with gold nanoparticles (AuNPs) encapsulating glucose oxidase (GOD) deposited on a gold electrode (Au) for glucose sensing, is described. Using the electrocatalytic synergy of AuNPs and GOD as a model of enzyme, the variation of the current (µA) as a function of the log of the glucose concentration (log [glucose]), shows three times higher sensitivity for the modified electrode (283.9) compared to that of the PEC/GOD modified electrode (93.7), with a detection limit of about 5 µM and a linearity range between 10 µM and 7 mM. The response of the PEC/AuNPs/GOD based biosensor also presents good reproducibility, stability, and negligible interfering effects from ascorbic acid, uric acid, urea, and creatinine. The applicability of the PEC/AuNPs/GOD based biosensor was tested in glucose-spiked saliva samples and acceptable recovery rates were obtained.

Voltammetric glucose biosensor based on glucose oxidase encapsulation in a chitosan-kappa-carrageenan polyelectrolyte complex.

In this work, a new design of voltammetric glucose biosensor, based on the encapsulation of glucose oxidase (GOx) in a chitosan/κ­carrageenan (CHIT/CAR) polyelectrolyte complex (PEC) using a simple coacervation process is presented. A conductometric monitoring of this is performed. Spectroscopic and morphological characterization of the PEC film encapsulating GOx is carried out. Compared to biosensors based on a chitosan film, a more sensitive voltammetric detection of glucose is obtained. Using square wave voltammetry (SWV), the CHIT/CAR PEC based biosensor exhibits a wide linearity range from 5 µM to 7 mM glucose with a detection limit of 5 µM. Excellent selectivity against ascorbic acid, uric acid and urea is observed and the applicability of the biosensor for glucose detection in spiked saliva samples was demonstrated.

Aflatoxin B1 Detection Using a Highly-Sensitive Molecularly-Imprinted Electrochemical Sensor Based on an Electropolymerized Metal Organic Framework.

A sensitive electrochemical molecularly-imprinted sensor was developed for the detection of aflatoxin B1 (AFB1), by electropolymerization of p-aminothiophenol-functionalized gold nanoparticles in the presence of AFB1 as a template molecule. The extraction of the template leads to the formation of cavities that are able to specifically recognize and bind AFB1 through π-π interactions between AFB1 molecules and aniline moities. The performance of the developed sensor for the detection of AFB1 was investigated by linear sweep voltammetry using a hexacyanoferrate/hexacyanoferrite solution as a redox probe, the electron transfer rate increasing when the concentration of AFB1 increases, due to a p-doping effect. The molecularly-imprinted sensor exhibits a broad linear range, between 3.2 fM and 3.2 µM, and a quantification limit of 3 fM. Compared to the non-imprinted sensor, the imprinting factor was found to be 10. Selectivity studies were also performed towards the binding of other aflatoxins and ochratoxin A, proving good selectivity.

Anticancer drug detection using a highly sensitive molecularly imprinted electrochemical sensor based on an electropolymerized microporous metal organic framework.

A simple and highly sensitive approach for the detection of the anti-neoplastic drug gemcitabine is presented, based on a one-step electropolymerized molecularly imprinted microporous-metal-organic-framework. The sensitive layer was prepared by electropolymerization of the aniline moieties of p-aminothiophenol- gold nanoparticles on the surface of gold electrodes tethered with p-aminothiophenol, in the presence of gemcitabine as a template molecule. Experimental parameters that control the performance of the sensor were investigated and optimized. Under optimal conditions a calibration curve was obtained in the linear range from 3.8 fM to 38 nM with a limit of detection of 3 fM. The obtained imprinted sensor has the advantages of easy manufacture, high sensitivity and selectivity and good reproducibility. Furthermore the feasibility of the proposed technique has been investigated on spiked serum samples and infusion solution containing gemcitabine.

Leakage interferences applied to surface plasmon analysis.

We report the experimental combination of leakage radiation microscopy with a Young slit experiment to address the spatial coherence properties of surface waves. We applied this method to measurements of surface plasmon polaritons (SPPs). The relationship between the spatial decay and interference contrast allows us to extract the degree of coherence. In a second step, we investigate the coherence properties of the plasmon in the weak coupling regime between fluorophores and metallic surfaces. Finally, a method is proposed to extract the propagation length of SPPs in a large variety of systems.

Formation and optical properties of silver perfluorodecanethiolate nanoparticles.

This article reports a new catalytic method for preparing nanoparticles of silver thiolate from silver nanoparticles scattered on a ZrO2-coated substrate. Such nanoparticles transform into silver (perfluoro) decanethiolate after immersion in a solution of (perfluoro) decanethiol in heptane. These transformations occur at room temperature and are catalysed by ZrO2. The silver decanethiolate is obtained as lamellar crystals while the silver perfluorodecanethiolate is obtained in amorphous state. The modifications of the sample optical properties due to this latter compound are studied in correlation with its surface morphology, according to different preparation conditions. It is shown that an antireflective effect in addition to the damping of the plasmon band of the silver nanoparticles can be responsible for a large transmittance enhancement in the near-UV and visible ranges. These effects are modulated by the possible oxidation of the silver nanoparticle surface. In the absence of silver oxidation, the silver perfluorodecanethiolate is obtained as contiguous spheroidal nanoparticles, while, in the presence of silver oxidation, this compound is mainly obtained as entangled nanowires.

Chemical growth and photochromism of silver nanoparticles into a mesoporous titania template.

Elaboration of mesoporous titanium oxide film supporting silver nanoparticles is described. Mesoporous titanium oxide films are characterized by TEM analysis. Titania films are infiltrated with a silver salt solution and chemical reduction treatments are performed using either a NaBH(4) or a formaldehyde solution. Infiltrated films are then characterized by TEM, SEM, AFM, UV-visible spectroscopy, X-ray diffraction, and Rutherford Backscattering Spectrometry (RBS). The utilization of a mesoporous titania substrate allows to control the nanoparticle size and the interparticle distance. RBS experiments provide the evidence that NaBH(4) treatment induces a strong accumulation of silver nanoparticles in the subsurface of the layer, while formaldehyde treatment induces the formation of silver nanoparticles embedded into almost the whole depth of the titania film. Large silver nanocrystals are also formed at the film surface whatever the reducer used. A broad visible absorption band related to the surface plasmon resonance (SPR) is obtained in both cases and is strongly red-shifted compared to the SPR obtained for silver nanoparticles inside a silica matrix. Moreover, irradiation with visible light causes the photooxidation of silver nanoparticles by titania and a complete discoloration of the material. The photooxidation is related to a drastic decrease in the silver nanoparticle size and is found to be reversible, particularly in the case of the material obtained by the formaldehyde reduction.

Validation of a conductometric bienzyme biosensor for the detection of proteins as marker of organic matter in river samples.

This article describes a conductometric bi-layer based bienzyme biosensor for the detection of proteins as a marker of organic matter in rivers. Proteins were chosen to be used as indicators of urban pollution. The working mechanism of the bienzyme biosensor is based on the enzymatic hydrolysis of proteins into several fractions (peptides and amino acids), which results in a local conductivity change depending of the concentration of proteins. In this work, we began with the optimization of biosensor response using bovine serum albumin (BSA) as standard protein. For this objective seven enzymatic biosensors were prepared: four enzymatic sensors with only one layer of enzyme (proteinase K, trypsin, pronase or protease X) and three other enzymatic sensors with two layers (first layer: membrane containing proteinase K; second layer: one of the three other enzymes: trypsin, pronase or protease X). The biosensors were obtained through the deposition of enzymatic layers and the cross-linking process between enzymes and BSA in saturated glutaraldehyde vapour. The response of the various biosensors, described previously, were compared with the values of total organic carbon (TOC), and those of organic nitrogen (Norg), as determined by the laboratory accredits (CEMAGREF of Lyon) using the traditional method of analysis (NF EN 1484, infrared spectroscopy) and (NF EN 25663, mineralization/colorimetry assay) respectively for each water sample obtained from different sites in Lyon (France). The linear correlations obtained with the response of the seven biosensors showed the most important indices of correlations for the biosensor with two enzymatic layers: proteinase K + pronase (pkp). The optimum conditions for the preparation of the pkp biosensor increased the sensitivity and gave a limit of quantification of 0.583 microg/L for TOC and 0.218 microg/L for Norg in water samples. This sensor shows good reproducibility (2.28%), a capacity to be used at temperatures range 10-30 degrees C (depending on the season) and moreover a long lifetime (5 weeks).

A novel nitrite biosensor based on conductometric electrode modified with cytochrome c nitrite reductase composite membrane.

A conductometric biosensor for nitrite detection was developed using cytochrome c nitrite reductase (ccNiR) extracted from Desulfovibrio desulfuricans ATCC 27774 cells immobilized on a planar interdigitated electrode by cross-linking with saturated glutaraldehyde (GA) vapour in the presence of bovine serum albumin, methyl viologen (MV), Nafion, and glycerol. The configuration parameters for this biosensor, including the enzyme concentration, ccNiR/BSA ratio, MV concentration, and Nafion concentration, were optimized. Various experimental parameters, such as sodium dithionite added, working buffer solution, and temperature, were investigated with regard to their effect on the conductance response of the biosensor to nitrite. Under the optimum conditions at room temperature (about 25 degrees C), the conductometric biosensor showed a fast response to nitrite (about 10s) with a linear range of 0.2-120 microM, a sensitivity of 0.194 microS/microM [NO(2)(-)], and a detection limit of 0.05 microM. The biosensor also showed satisfactory reproducibility (relative standard deviation of 6%, n=5). The apparent Michaelis-Menten constant (K(M,app)) was 338 microM. When stored in potassium phosphate buffer (100mM, pH 7.6) at 4 degrees C, the biosensor showed good stability over 1 month. No obvious interference from other ionic species familiar in natural waters was detected. The application experiments show that the biosensor is suitable for use in real water samples.

Development of a conductometric phosphate biosensor based on tri-layer maltose phosphorylase composite films.

A conductometric biosensor for phosphate detection was developed using maltose phosphorylase (MP) from recombinant Escherichia coli immobilized on a planar interdigitated electrode by cross-linking with saturated glutaraldehyde (GA) vapour in the presence of bovine serum albumin (BSA). The process parameters for the fabrication of the mono-enzymatic sensor and various experimental variables such as the enzyme loading, time of immobilization in saturated GA vapour, working buffer solution and temperature were investigated with regard to their influence on sensitivity, detection limit, dynamic range, operational and storage stability. The biosensor can work well at the temperature between 20 degrees C and 50 degrees C, and reach 90% of steady-state conductance in about 10s. The sensor has two linear ranges, one is from 1.0 microM to 20 microM phosphate with a detection limit of 1.0 microM, and the other is between 20 microM and 400 microM phosphate. When stored in citrate buffer (0.1M, pH 6.0) at 4 degrees C, the biosensor showed good stability over two months. No obvious interference from other anionic species like SO(4)(2-), Cl(-), NO(3)(-), NO(2)(-) and HCO(3)(-) was detected. The biosensor is suitable for use in real water samples.

Evaluation of endothelial cell adhesion onto different protein/gold electrodes by EIS.

To study cell attachment to biomaterials, several proteins such as fibronectin, collagen IV, heparin, immunoglobulin G, and albumin have been deposited onto polystyrene adsorbed on a self-assembled monolayer (silane or thiol) on glass or gold, respectively. The different steps of this multilayer assembly have been characterized by electrochemical impedance spectroscopy (EIS). These data are compared to those of adhesion rate, viability percentage, and cytoskeleton labeling for a better understanding of the cell adhesion process to each protein. All the proteins are endothelial cell adhering biomolecules but not with the same features. A linear relationship has been established between adhesion rate and resistance of the endothelial cell/protein interface for all negatively charged proteins.