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The ecotoxicological impact of leptospermone, a β-triketone bioherbicide, on the bacterial community of two arable soils was investigated. Soil microcosms were exposed or not (control, 0×) to leptospermone applied at 1× or 10× recommended dose. Leptospermone was moderately adsorbed to both soils (i.e.: Kfa  1.2 and Koc  140 ml.g-1). Dissipation of leptospermone was lower in sterilized than in unsterilized soils suggesting that it was mainly influenced by biotic factors. Within 45 days, leptospermone disappeared almost entirely from one of the two soils (i.e. DT50 < 10 days), while 25% of the leptospermone remained in the other. The composition of the microbial community assessed by qPCR targeting 11 microbial groups was found to be significantly modified in soil microcosms exposed to leptospermone. Pyrosequencing of 16S rRNA gene amplicons showed a shift in the bacterial community structure and a significant impact of leptospermone on the diversity of the soil bacterial community. Changes in the composition as well as in the α- and β-diversity of microbial community were transient in the soil able to fully dissipate the leptospermone. They were persistent in the soil where leptospermone remained. To conclude the bacterial community of the two soils was sensitive to leptospermone and its resilience was observed only when leptospermone was fully dissipated.

International audience; A novel dual electrochemical immunosensor was fabricated for the rapid and sensitive detection of enrofloxacin (EF) antibiotic in meat. Anti-quinolone antibody was immobilized onto screen-printed dual carbon electrodes via carbodiimide coupling. A new electrochemical probe was synthesized by conjugating difloxacin and aminoferrocene, whose oxidation was measured at +0.2 V vs. Ag/AgCl by differential pulse voltammetry. The detection principle was based on the competitive binding of this conjugate and free EF on immobilized antibodies. The proposed immunosensor allowed detection of EF at concentrations ranging from 0.005 µg.mL-1 to 0.01 µg.mL-1 with a detection limit of 0.003 µg.mL-1. The immunosensor was stable for at least 1 month at 4°C and displayed a good specificity for other fluoroquinolones. The new dual electrode design offered an improved accuracy as one electrode was used as negative control. The efficiency of the sensor and the adequacy of the extraction process were finally validated by detecting EF in different meat samples.

International audience; The bis-triazole ligand and its corresponding copper complexes were synthesized and characterized for the first time and proposed as new labels for the development of electrochemical aptasensors. The bis-triazole ligand was prepared from methyl 1,6-heptadiyne-4-carboxylate and 2-(azidomethyl)phenol using classical CuAAC in presence of different copper salts. The X-ray structure of bis-triazole showed a symmetry center (C1). UV-Vis and X-band EPR spectra showed that the coordination capacity of the bis-triazole ligand was improved in the presence of triethylamine due to deprotonation of the triazole and phenolate moieties. After complexation with copper, the obtained complex was successfully attached to an anti-estradiol aptamer through thiol-maleimide coupling, and the resulting labelled aptamer was immobilized on a carbon screen-printed electrode by carbodiimide coupling. The electrochemical response of the resulting sensor was shown to decrease in the presence of estradiol, demonstrating that the developed complexes can be applied for the development of aptasensors.

BACKGROUND: The occurrence of pharmaceutical compounds in wastewater has become a major concern for human health and the environment. Therefore, it is an important challenge to improve conventional wastewater treatment to remove these compounds. Coupling biological treatment with advanced oxidation technology has been widely studied in the literature, but only sequential associations of the two processes have been used. This study proposes an innovative concept based on a real integration of the photocatalytic oxidation process in a continuous recycling loop on a membrane bioreactor. The role of the oxidation here is not to completely degrade pharmaceuticals, but instead to oxidize them moderately to increase their biodegradability so that they can be eliminated by the biological process. RESULTS: Preliminary experiments on the oxidation process indicated that a flux density of 5 W m−2 was sufficient to increase biodegradability and decrease the toxicity of a cocktail of three pharmaceuticals. Then the performance of a 20-L continuous membrane bioreactor treating wastewater with seven pharmaceuticals, without and with pre-oxidation at 5 W m−2, were compared. Pre-oxidation increased the global removal for some recalcitrant pharmaceuticals (from 3% to 47% for diclofenac and from 1% to 44% for furosemide) without affecting either the removal of carbon, nitrogen or phosphorous by activated sludge, or of already highly removed pharmaceuticals. CONCLUSION: This work proves the feasibility of an innovative concept of a continuous hybrid process coupling a photocatalytic oxidation process and a membrane bioreactor for the treatment of pharmaceuticals in wastewater, with low cost and size. © 2020 Society of Chemical Industry (SCI)., This work was supported by the Program ‘SUDOE’ on the project Innovec'EAU 2016-2019 under award number SUDOE SOE1/P1/F0173. We thank R. Garcia and J. J. Huc for their help in designing and building the demonstrator, and Victor Pueyo for doing the analysis.

Maize cultivators often use β-triketone herbicides to prevent the growth of weeds in their fields. These herbicides target the 4-HPPD enzyme of dicotyledons. This enzyme, encoded by the hppd gene, is widespread among all living organisms including soil bacteria, which are considered as “non-target organisms” by the legislation. Within the framework of the pesticide registration process, the ecotoxicological impact of herbicides on soil microorganisms is solely based on carbon and nitrogen mineralization tests. In this study, we used more extensive approaches to assess with a lab-to-field experiment the risk of β-triketone on the abundance and the diversity of both total and hppd soil bacterial communities. Soil microcosms were exposed, under lab conditions, to 1× or 10× the recommended dose of sulcotrione or its commercial product, Decano®. Whatever the treatment applied, sulcotrione was fully dissipated from soil after 42 days post-treatment. The abundance and the diversity of both the total and the hppd bacterial communities were not affected by the herbicide treatments all along the experiment. Same measurements were led in real agronomical conditions, on three different fields located in the same area cropped with maize: one not exposed to any plant protection products, another one exposed to a series of plant protection products (PPPs) comprising mesotrione, and a last one exposed to different PPPs including mesotrione and tembotrione, two β-triketones. In this latter, the abundance of the hppd community varied over time. The diversity of the total and the hppd communities evolved over time independently from the treatment received. Only slight but significant transient effects on the abundance of the hppd community in one of the tested soil were observed. Our results showed that tested β-triketones have no visible impact toward both total and hppd soil bacteria communities.

Metallic complexes of macrocycles chelators 1,4,7-triazacyclononane-N,N,N-triacetic acid (NOTA) and 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) were synthetized with iron (III) giving Fe(III)-DOTA and Fe(III)-NOTA complexes. They were studied in comparison of ferricyanide and ferrocenemethanol on cyclic voltammetry with glassy carbon working electrode (GC) and screen-printed carbon electrode (SPCE). Diffusion coefficients and heterogeneous electron transfer rate constants were determined with Randles-Sevcik and Nicholson-Lavagnini methods. Using SPCE. The average values of diffusion coefficient and transfer rate constant were respectively of 1.34 × 10−6 cm2 s−1 and 1.01 × 10−3 cm s−1 for Fe(III)-DOTA, and 4.32 × 10−6 cm2 s−1 and 1.14 × 10−3 cm s−1 for Fe(III)-NOTA. Interestingly, characteristics of the synthetized complexes on SPCE compete advantageously with those of ferricyanide and ferrocenemethanol, making Fe(III)-DOTA and Fe(III)-NOTA potential markers for future biosensors development.

With the increasing incidence and prevalence of antibiotic contamination in animal-derived food and drug resistance around the world, early and specific detection of antibiotic residues has garnered significant attention. Herein, for the first time, we devise a facile, label free and portable aptasensor for quantitative determination of Kanamycin (KANA) by electrochemical impedance spectroscopy (EIS), which accompanying the assembly of in-vitro selected single strand DNA (ssDNA) anti-KANA-aptamer functionalized screen printed carbon electrodes (SPCEs) used as transducer. The target detection is based on specific recognition by KANA-aptamer covalently immobilized on SPCEs surface. The surface morphology and electrochemical properties of aptasensor were characterized using Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), atomic force microscopy (AFM), cyclic voltammetry (CV) and EIS. Under optimized experimental conditions, the devised aptasensor exhibited a dynamic range 1.2–600 ng mL−1 with linearity 1.2–75 ng mL−1 and limit of detection (LOD) 0.11 ng mL−1 (S/N = 3) of KANA. The developed aptasensor are endowed with a good selectivity and specificity to KANA without interference from competitive analogues (streptomycin and gentamicin). For practical application, the aptasensor performance was verified in spiked milk samples and the acceptable recovery percentage of 96.88–100.5% (%RSD = 4.56, n = 3) was obtained in milk samples.

The paper describes the development of an amperometric biosensor using Prussian Blue (PB) modified electrodes containing xanthine oxidase (XOD). The enzyme is immobilized by photo-polymerization into an azide-unit pendant water-soluble photopolymer (PVA-AWP). The parameters of the fabrication of the biosensor, XOD:PVA/AWP ratio, crosslinking irradiation time, and XOD charge, were optimized. Operational conditions for electrode preparation were defined as 1:2 ratio of XOD:PVA/AWP; exposure time to neon light of 30 min; pH = 7.5 at room temperature and enzymatic charge of 8 mU per electrode. The biosensors showed stable, fast, simple, selective, cost-effective and sensitive (-2.72E-8 A mol L-1), with a good linear range (1.0–75 μmol L-1), and respectively detection and quantification limits for antioxidants of 2.17, and 7.15 μmol L-1. The applicability of this biosensor was demonstrated by in vitro analysis of gallic acid as standard antioxidant and Amazonian fruits as natural sources.

International audience; This work presents the synthesis and characterization of salen/salan metal complexes for their future application as electrochemicallabels in affinity sensors. Due to its stability and electrochemical properties, an oxovanadium salan complex was selected andcoupled to an estradiol-specific aptamer. The response of the resulting aptasensor was shown to decrease with increasing estradiolconcentration.

RNA represents an extremely promising and yet challenging therapeutic target. Here, we report the design of a series of C-nucleosides as original RNA binders. Some of them bind strongly and selectively to A-site prokaryotic ribosomal RNA.

A quenching based apta-sensing platform was developed for the detection of Patulin. Three different aptamer sequences were studied to screen the aptamer with the maximum affinity towards Patulin. Carboxyfluorescein (CFL) was used as a fluorescent dye while –COOH functionalized multiwall carbon nanotubes (MWCNTs) were applied as novel nanoquenchers. Aptamer tagged at the 3′ end with 40 nucleotide bases exhibited the maximum affinity towards Patulin and caused substantial fluorescence recovery. Interestingly, the limit of detection (LOD) and limit of quantification (LOQ) were calculated as 0.13 μg L−1 and 0.41 μg L−1 respectively. Commonly occurring mycotoxins in food were also tested to confirm the selectivity of apta-assay. The developed apta-assay was applied to a spiked apple juice sample and toxin recoveries were observed ranging from 96% to 98% (n = 3). These results demonstrated the potential of the developed apta-assay for the selective detection and quantification of Patulin in food samples.

International audience; Biological and synthetic recognition elements are at the heart of the majority of modern bioreceptor assays. Traditionally, enzymes and antibodies have been integrated in the biosensor designs as a popular choice for the detection of toxin molecules. But since 1970s, alternative biological and synthetic binders have been emerged as a promising alternative to conventional biorecognition elements in detection systems for laboratory and field-based applications. Recent research has witnessed immense interest in the use of recombinant enzymatic methodologies and nanozymes to circumvent the drawbacks associated with natural enzymes. In the area of antibody production, technologies based on the modification of in vivo synthesized materials and in vitro approaches with development of “display “systems have been introduced in the recent years. Subsequently, molecularly-imprinted polymers and Peptide nucleic acid (PNAs) were developed as an attractive receptor with applications in the area of sample preparation and detection systems. In this article, we discuss all alternatives to conventional biomolecules employed in the detection of various toxin molecules We review recent developments in modified enzymes, nanozymes, nanobodies, aptamers, peptides, protein scaffolds and DNazymes. With the advent of nanostructures and new interface materials, these recognition elements will be major players in future biosensor development.

International audience; Ochratoxin A (OTA) is a carcinogenic mycotoxin that contaminates food such as cereals, wine and beer; therefore it represents a risk for human health. Consequently, the allowed concentration of OTA in food is regulated by governmental organizations and its detection is of major agronomical interest. In the current study we report the development of an electrochemical aptasensor able to directly detect trace OTA without any amplification procedure. This aptasensor was constructed by coating the surface of a gold electrode with a film layer of modified polypyrrole (PPy), which was thereafter covalently bound to polyamidoamine dendrimers of the fourth generation (PAMAM G4). Finally, DNA aptamers that specifically binds OTA were covalently bound to the PAMAM G4 providing the aptasensor, which was characterized by using both Atomic Force Microscopy (AFM) and Surface Plasmon Resonance (SPR) techniques. The study of OTA detection by the constructed electrochemical aptasensor was performed using Electrochemical Impedance Spectroscopy (EIS) and revealed that the presence of OTA led to the modification of the electrical properties of the PPy layer. These modifications could be assigned to conformational changes in the folding of the aptamers upon specific binding of OTA. The aptasensor had a dynamic range of up to 5 μg L−1 of OTA and a detection limit of 2 ng L−1 of OTA, which is below the OTA concentration allowed in food by the European regulations. The efficient detection of OTA by this electrochemical aptasensor provides an unforeseen platform that could be used for the detection of various small molecules through specific aptamer association.

Ochratoxin A (OTA), a widespread food contaminant, is considered as one of the most hazardous mycotoxins commonly occurring in beer. Herein, we describe a novel impedimetric aptasensor based on Pencil Graphite Electrodes (PGE) for the analysis of OTA. Taking advantage of its affordability and high active surface area, a PGE was modified with a specific aptamer, to achieve sensitive detection of OTA. This strategy relies on the target-induced increase in the electron transfer resistance (Rct) upon OTA recognition using ferro/ferricyanide as a redox probe. With the addition of OTA, the formation of OTA–aptamer complex resulted in an increase of Rct. The change of Rct strongly depends on OTA concentration, which is applied for mycotoxin quantification. An acceptable linear range was obtained from 0.1 to 2.0 ng mL−1 with a very good limit of detection as low as 0,1 ng mL−1. This simple-to-design aptasensor was successfully tested in spiked beer samples, with OTA concentrations in the range of 0.4–1.6 ng mL−1, attesting a recovery percentage of 93.4 ± 6.6%. It also showed a high level of stability and reproducibility thanks to the electro-grafting method via in-situ generation of 4-carboxyphenyl diazonium salts. This PGE-based platform is a versatile approach for the design of low-cost aptasensors for food safety control.

Electrochemical molecularly imprinted polymers (e-MIPs) were for the first time introduced in screen-printed carbon electrodes (SPCE) as the sensing element for the detection of an organic pollutant. To play this sensing role, a redox tracer was incorporated inside the binding cavities of a cross-linked MIP, as a functional monomer during the synthesis step. Ferrocenylmethyl methacrylate was used for this purpose. It was associated with 4-vinylpyridine as a co-functional monomer and ethylene glycol dimethacrylate as cross-linker for the recognition of the endocrine disruptor, Bisphenol A (BPA), as a target. Microbeads of e-MIP and e-NIP (corresponding non-imprinted polymer) were obtained via precipitation polymerization in acetonitrile. The presence of ferrocene inside the polymers was assessed via FTIR and elemental analysis and the polymers microstructure was characterized by SEM and nitrogen adsorption/desorption experiments. Binding isotherms and batch selectivity experiments evidenced the presence of binding cavities inside the e-MIP and its high affinity for BPA compared to carbamazepine and ketoprofen. e-MIP (and e-NIP) microbeads were then incorporated in a graphite-hydroxyethylcellulose composite paste to prepare SPCE. Electrochemical properties of e-MIP-SPCE revealed a high sensitivity in the presence of BPA in aqueous medium compared to e-NIP-SPCE with a limit of detection (LOD) of 0.06 nM. Selectivity towards carbamazepine and ketoprofen was also observed with the e-MIP-SPCE.

International audience; This paper presents for the first time the integration of a biosensor for the on-line, real-time monitoring of a photocatalytic degradation process. Paracetamol was used as a model molecule due to its wide use and occurrence in environmental waters. The biosensor was developed based on tyrosinase immobilization in a polyvinylalcohol photocrosslinkable polymer. It was inserted in a computer-controlled flow system installed besides a photocatalytic reactor including titanium dioxide (TiO2) as photocatalyst. It was shown that the biosensor was able to accurately monitor the paracetamol degradation with time. Compared with conventional HPLC analysis, the described device provides a real-time information on the reaction advancement, allowing a better control of the photodegradation process.

International audience; The fate of benzo(a)pyrene (BaP), a ubiquitous contaminant reported to be persistent in the environment, is largely controlled by its interactions with the soil organic matter. In the present study, the spectral characteristics of fluorophores present in the physical fractions of the soil organic matter were investigated in the presence of pure BaP solution. After extraction of humic substances (HSs), and their fractionation into fluvic acid (FA) and humic acid (HA), two fluorescent compounds (C1 and C2) were identified and characterized in each physical soil fraction, by means of fluorescence excitation-emission matrices (FEEMs) and Parallel Factor Analysis (PARAFAC). Then, to each type of fraction having similar DOC content, was added an increasing volume of pure BaP solution in attempt to assess the behavior of BaP with the fluorophores present in each one. The application of FEEMs-PARAFAC method validated a three-component model that consisted of the two resulted fluorophores from HSs, FA and HA (C1 and C2) and a BaP-like fluorophore (C3). Spectral modifications were noted for components C2HSs (C2 in humic substances fraction) (λex/λem: 420/490-520 nm), C2FA (C2 in fulvic acid fraction) (λex/λem: 400/487(517) nm) and C1HA (C1 in humic acid fraction) (λex/λem: 350/452(520) nm). We explored the impact of increasing the volume of the added pure BaP solution on the scores of the fluorophores present in the soil fractions. It was found that the scores of C2HSs, C2FA, and C1HA increased when the volume of the added pure BaP solution increased. Superposition of the excitation spectra of these fluorophores with the emission spectrum of BaP showed significant overlaps that might explain the observed interactions between BaP and the fluorescent compounds present in SOM physical fractions.

International audience; In this paper, we have reported a novel electrochemical aptamer–antibody based sandwich biosensor for the detection of lysozyme. In the sensing strategy, an anti-lysozyme aptamer was immobilized onto the carbon electrode surface by covalent binding via diazonium salt chemistry. After incubating with a target protein (lysozyme), a biotinylated antibody was used to complete the sandwich format. The subsequent additions of avidin-alkaline phosphatase as an enzyme label, and a 1-naphthyl phosphate substrate (1-NPP) allowed us to determine the concentration of lysozyme (Lys) via Differential Pulse Voltammetry (DPV) of the generated enzyme reaction product, 1-naphthol. Using this strategy, a wide detection range from 5 fM to 5 nM was obtained for a target lysozyme, with a detection limit of 4.3 fM. The control experiments were carried out by using bovine serum albumin (BSA), cytochrome c and casein. The results showed that the proposed biosensor had good specificity, stability and reproducibility for lysozyme analysis. In addition, the biosensor was applied for detecting lysozyme in spiked wine samples, and very good recovery rates were obtained in the range from 95.2 to 102.0% for lysozyme detection. This implies that the proposed sandwich biosensor is a promising analytical tool for the analysis of lysozyme in real samples.

International audience; Molecularly imprinted polymers (MIPs) for phosmet insecticide were synthesized by batch polymerization. The affinity of functional monomers to phosmet was tested using an original method involving an electrochemical biosensor based on acetylcholinesterase inhibition. It was demonstrated that association of phosmet with appropriate functional monomers resulted in a decrease of enzyme inhibition. Using this method, it was shown that N,N-methylenebis(acrylamide) displayed the highest interactions with phosmet using DMSO as solvent. These results were in good accordance with those obtained by conventional computational modeling. Molecularly imprinted polymers (MIPs) and non-imprinted polymers (NIPs) were synthesized and adsorption isotherms were studied to describe their interaction with phosmet. Freundlich isotherm was able to fit phosmet adsorption on MIPs with good agreement (R2 = 0.9). The pre-exponential factor KF determined for MIPs was 1.439mg(1-N)g-1LN, more that 10 times higher than for NIPs (0.125mg(1-N)g-1. LN), indicating an increase of binding sites number and average affinity.

International audience; This paper describes for the first time the selection of aptamers selective to penicillin. Aptamers were selected using a specific process called Capture-SELEX (Systematic Evolution of Ligands by Exponential Enrichment). This technique is based on the selection of DNA aptamers using penicillin G in solution while the ssDNA library is fixed on a support. One aptamer showing a good affinity to penicillin was finally selected and tested in electrochemical sensor configuration, using electrochemical impedance spectroscopy as detection technique. The developed aptasensor allowed the detection of penicillin in a wide concentration range, comprised between 0.4 and 1000µgL-1 Such performance was compatible with milk analysis, as the maximum residue limit tolerated in this matrix is 4µgL-1. The selectivity of the developed sensor was also studied, showing that the sensor was also able to bind other beta-lactam antibiotics, although with a weaker affinity. Finally the sensor was used for detection of penicillin G in milk. It was shown that a simple sample treatment with isopropanol followed by filtration was sufficient to eliminate matrix effects, allowing the determination of penicillin in milk at concentrations compatible with legislation requirements.

This study presents a novel approach for assessing the risk of agrochemicals in soil microcosms through the use of non-targeted metabolomics. The metabolome of treated soils was extracted and tested through LCMS profiling in order to generate an "Environmental Metabolic Footprint" (EMF). A dynamic characterization of pollution biomarkers was obtained through a multivariate statistical analysis of EMF data, where our results show the possible evolution towards a state of resilience. The EMF methodology was applied to two β-triketone herbicides in soil microcosms: one natural, leptospermone, and one synthetic, sulcotrione. In spite of a four-fold higher application dose, leptospermone exhibited a lower resilience time than did sulcotrione (ca. 30 days vs ca. 45 days respectively).

International audience; In this study, a simple TAMRA (tetramethyl-6-carboxyrhodamine) quenching-based aptasensing platform was designed for the detection of aflatoxin B1 (AFB1). Here, we compared the analytical performance of two aptamer sequences: seqA and seqB. The AFB1 detection was based on the interactions of FAM (carboxyfluorescein)-labeled aptamer with TAMRA-labeled DNA complementary strand in the presence and absence of target analyte. Under optimized experimental conditions, TAMRA-labeled strand quenched the fluorescence response of FAM-labeled aptamer due to the noncovalent interaction between the two DNA strands. The binding of AFB1 induced the complex formation and weakened the interaction between FAM-labeled aptamer and TAMRA-labeled complementary strand, resulting in the fluorescence recovery. By using this principle concept, an assay was constructed for the detection of AFB1. The method exhibited good sensitivity, good selectivity with a limit of detection of 0.2 ng ml−1, and a wide linear range from 0.25 to 32 ng ml−1. For real sample application, the aptasensors were tested in beer and wine samples, with good recovery rates obtained for AFB1 detection.

International audience; A novel PCR-free DNA-based assay was developed for the detection of Vibrio spp. A sandwich hybridization format using an immobilized capture probe and a labeled signal probe was selected and combined with chemiluminescent method for the detection of the RNA target. In a first step, probes were validated using positive controls (PCs). A linearity was observed between 0.1 and 2.5 nM of PC, and detection limit was determined as 0.1 nM. In a second step, specificity was checked by using RNA extracted from a panel of 31 environmental bacterial strains. Detection limit of 5 ng μL−1 of total fragmented RNA was obtained, and the assay allowed a good discrimination between the 21 Vibrio and the 10 non-Vibrio strains tested. Finally, the DNA-based assay was successfully applied to analysis of spiked and natural environmental samples. Stability and analysis time of the DNA-based assay were also investigated to optimize working conditions. We demonstrated that microplates can be coated beforehand with capture probe and stored at 4 °C without any buffer in wells for at least 30 days. The use of the pre-made plates enables the assay to be completed in 2 h. The developed assay appeared as an interesting tool to determine the presence of bacteria in environmental samples

International audience; An ultra-sensitive screen-printed biosensor was successfully developed for phosmet detection in olive oil, based on a genetically-engineered acetylcholinesterase (AChE) immobilized in a azide-unit water-pendant polyvinyl alcohol (PVA-AWP)/Fe-Ni alloy nanocomposite. Fe-Ni not only allowed amplifying the response current but also lowering the applied potential from 80 mV to 30 mV vs Ag/AgCl. The biosensor showed a very good analytical performance for phosmet detection, with a detection limit of 0.1 nM. This detection limit is lower than the allowable concentrations set by international regulations. In addition to the good reproducibility, operational and storage stability, the developed biosensor was successfully used for the determination of phosmet in olive oil samples without any laborious pre-treatment. The phosmet recovery rate was about 96% after a simple liquid-liquid extraction.

International audience; A Surface Plasmon Resonance (SPR) sensor for the quantitation of lysozyme dimer in monomer-dimer mixtures, reaching a detection limit of 1.4 nM dimer, has been developed. The sensor is based on an aptamer which, although developed for the monomeric form, binds also the dimeric form but with a strikingly different kinetics. The aptasensor was calibrated using a dimer obtained by cross-linking. Sensorgrams acquired with the aptasensor in monomer-dimer mixtures were analysed using Principal Components Analysis and Multiple Regression to establish correlations with the dimer content in the mixtures. The method allows the detection of 0.1–1% dimer in monomer solutions without any separation. As an application, the aptasensor was used to qualitatively observe the initial stages of aggregation of lysozyme solutions at 60 °C and pH 2, through the variations in lysozyme dimer amounts. Several other methods were used to characterize the lysozyme dimer obtained by cross-linking and confirm the SPR results. This work highlights the versatility of the aptasensor, which can be used, by simply tuning the experimental conditions, for the sensitive detection of either the monomer or the dimer and for the observation of the aggregation process of lysozyme.

A novel strategy based on direct electrochemistry of amino group on screen printed carbon electrode surface (SPCE) was purposed. The purposed method was employed for the label free detection of ochratoxin A (OTA). A long spacer arm of hexamethyldiamine was immobilized on SPCE via electrochemical oxidation of its terminal amino-group. The activated carboxy-aptamer was covalently linked to other amino terminal group of immobilized hexamethyldiamine. The designed immobilized macromolecules resulted in the formation of long clusters on SPCE surface, while aptamer acted as gate to block the entrance of these clusters. The aptamer gates were closed due to change in conformation of aptamer upon target analyte binding, decreasing the electrochemical signal. The decrease in electrochemical signal was used for the detection of target molecule.