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We report a sensor for phosphate detection based on screen-printed electrodes modified with carbon black nanoparticles. The phosphate was measured in amperometric mode via electrochemical reduction of molybdophosphate complex. Carbon black nanoparticles demonstrated the ability to quantify the molybdophosphate complex at a low applied potential. Some analytical parameters such as the working solution (sulfuric acid 0.1M), applied potential (0.125V vs Ag/AgCl), and molybdate concentration (1mM) were optimized. Using these conditions, a linear range of 0.5-100µM was observed with a detection limit of 0.1µM, calculated as three times the standard deviation of the blank divided by the slope of calibration curve. The system was challenged in drinking, river, aquarium, and waste water samples yielding satisfactory recovery values in accordance with a spectrophotometric reference method which demonstrated the suitability of the screen-printed electrode modified with carbon black nanoparticles coupled with the use of molybdate to detect phosphate in water samples., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Trehalose is a disaccharide important in foods, serving as a glucose source in many and also as an additive in the food preparation. Because of its peculiar physico-chemical properties it plays an important role as preservative in drying and deep-freezing treatments. A new biosensor for trehalose determination has been realized by means of a flow system, based on a reactor in which the trehalase enzyme catalyses its hydrolysis into two alpha,d-glucose molecules, and a GOD (glucose oxidase) amperometric biosensor is employed for the glucose determination. The optimum operative conditions have been laid out and a particular attention has been paid to the immobilization procedure of the two enzymes. The electrode used is of the SPE (screen-printed electrode) type and has been activated with the Prussian Blue (PB) and then assembled using GOD immobilized with Nafion. The reactor has been prepared with the trehalase enzyme chemically immobilized on an Immunodyne ABC membrane. As demonstration of its utility, the biosensor has been tested on a real sample of Boletus edulis mushroom.

The production and assembling of disposable electrochemical AFM1 immunosensors, which can combine the high selectivity of immunoanalysis with the ease of the electrochemical probes, has been carried out. Firstly immunoassay parameters such as amounts of antibody and labelled antigen, buffer and pH, length of time and temperature of each steps (precoating, coating, binding and competition steps) were evaluated and optimised in order to set up a spectrophotometric enzyme-linked immunosorbent assay (ELISA) procedure. This assay exhibited a working range between 30 and 160 ppt in a direct competitive format. Then electrochemical immunosensors were fabricated by immobilising the antibodies directly on the surface of screen-printed electrodes (SPEs), and allowing the competition to occur between free AFM1 and that conjugated with peroxidase (HRP) enzyme. The electrochemical technique chosen was the chronoamperometry, performed at -100 mV. Furthermore, studies of interference and matrix effects have been performed to evaluate the suitability of the developed immunosensors for the analysis of aflatoxin M1 directly in milk. Results have shown that using screen-printed electrodes aflatoxin M1 can be measured with a detection limit of 25 ppt and with a working range between 30 and 160 ppt. A comparison between the spectrophotometric and electrochemical procedure showed that a better detection limit and shorter analysis time could be achieved using electrochemical detection.

Novel planar glucose biosensors to be used for continuous monitoring have been developed. The electrodes are produced with the "screen printing" technique, and present a high degree of reproducibility together with a low cost and the possibility of mass production. Prior to enzyme immobilisation, electrodes are chemically modified with ferric hexacyanoferrate (Prussian Blue). This allows the detection of the hydrogen peroxide produced by the enzymatic reaction catalysed by GOD, at low applied potential (ca. 0.0 V versus Ag/AgCl), highly limiting any electrochemical interferences. The layer of Prussian Blue (PB) showed a high stability at the working conditions (pH 7.4) and also after 1 year of storage dry at RT, no loss of activity was observed. The assembled glucose biosensors, showed high sensitivity towards glucose together with a long-term operational and storage stability. In a continuous flow system, with all the analytical parameters optimised, the glucose biosensors detected glucose concentration as low as 0.025 mM with a linear range up to 1.0mM. These probes were also tested over 50-60 h in a continuous flow mode to evaluate their operational stability. A 0.5 mM concentration of glucose was continuously fluxed into a biosensor wall-jet cell and the current due to the hydrogen peroxide reduction was continuously monitored. After 50-60 h, the drift of the signal observed was around 30%. Because of their high stability, these sensors suggest the possibility of using such biosensors, in conjunction with a microdialysis probe, for a continuous monitoring of glucose for clinical purposes.

The construction of an electrochemical immunosensor coupled to differential pulse voltammetry (DPV) for the detection of domoic acid (DA), a neurotoxic aminoacid responsible for the human syndrome known as "Amnesic Shellfish Poisoning" (ASP), is proposed here. The method involves the use of disposable screen-printed electrodes (SPEs) for the immunosensor development based on a "competitive indirect test". Domoic acid conjugated to bovine serum albumin (BSA-DA) was coated onto the working electrode of the SPE, followed by incubation with sample (or standard toxin) and anti-DA antibody. An anti-goat IgG-alkaline phosphatase (AP) conjugate was used for signal generation. A spectrophotometric enzyme-linked immunosorbent assay (ELISA) was used in a preliminary phase of development, prior to transferring the assay to the SPEs. Results showed a detection limit equal to 5 ng/ml of toxin. The electrochemical system is simple and cost-effective due to the disposable nature of the SPEs, and the analysis time is 150 min, shorter than that for the spectrophotometric method. The suitability of the assay for DA quantification in mussels was also evaluated. Samples were spiked with DA before and after the sample treatment to study the extraction efficiency and the matrix effect, respectively. After treatment, samples were analysed using a 1:250 v/v dilution in PBS-M (phosphate saline buffer pH 7.4 + CH3OH 10%) to minimise the matrix effect and allow for the detection of 20 microg/g of DA in mussel tissue. This represents the maximum acceptable limit defined by the Food and Drug Administration [Compliance Programme 7303.842. Guidance Levels, Table 3, p. 248, http://www.fda.org]. The optimised ELISA systems were then used, in parallel with a conventional HPLC method, to detect and confirm DA in shellfish extract in order to verify the performance of the electrochemical system. Very good recoveries were obtained, demonstrating the suitability of the proposed assay for accurate determination of the DA concentration in mussel samples.

Electropolymerisation of nonconducting polymer, poly-(1,2-diaminobenzene) on the top of Prussian Blue (PB) modified electrode led to significant improvement of resulting hydrogen peroxide transducer selectivity and operational stability. The reported transducer retained 100% of response during 20 h under the continuous flow of 0.1 mM H(2)O(2), and thus improves the stability level in selective peroxide detection by one order of magnitude. The selectivity value of the PB-poly(1,2-DAB) based H(2)O(2) sensor in relation to ascorbate is approximately 600. No signals to acetaminophen and urate were investigated. PB-poly(1,2-diaminobenzene) modified electrode allows the detection of H(2)O(2) in the flow-injection mode down to 10(-7) M with the sensitivity 0.3 A M(-1) cm(-2), which is only two times lower compared to the uncovered PB based transducer.

An amperometric biosensor for the determination of L-lactic acid in probiotic yogurts has been assembled using L-lactate dehydrogenase (EC 1.1.1.27, LDH) entrapped in 1% (v/v) neutralized Nafion® solution deposited on Variamine blue redox mediator modified screen-printed electrodes. The Variamine blue was previously covalently linked to oxidized single-walled carbon nanotubes and used for modifying screen-printed electrodes. The electrochemical cell, containing the L-lactate biosensor operating at an applied working potential of +200 mV vs. Ag|AgCl, was coupled with a microdialysis fiber and connected with a flow system, thus obtaining a microdialysis based sampling experimental set-up. Various analytical parameters, such as the cofactor concentration (2 mM, NAD+), the flow rate (10.5 μL/min), the applied working potential (+200 mV vs. Ag|AgCl), the working buffer (50 mM phosphate buffer +0.1 M KCl), and pH (7.5), were optimized in batch amperometric experiments. The dynamic linear working range was comprised between 2·10-4 and 1·10-3 M. The proposed biosensor was challenged with real samples of yogurt, properly diluted in working buffer, and the performances of the L-lactate biosensor were compared with a commercially available kit for the determination of L-lactic acid in foodstuffs from R-Biopharm GmbH, Germany, showing a good agreement. [ABSTRACT FROM AUTHOR]

An electrochemical biosensor to detect biogenic amines in saliva was developed. This biosensor is based on a Prussian Blue screen-printed electrode, which senses the H2O2 produced by the reaction catalyzed by the diamine oxidase enzyme. The probe was connected with a portable instrument and chronoamperometric drop measurements were performed. After evaluation of the biosensor's ability to interact with different amines, putrescine was chosen as the reference amine. A detection limit of 1 × 10-5 M and a working range between 2 × 10-5 and 3 × 10-4 M were obtained. Once optimized, the saliva treatment, a negligible matrix effect, and an average recovery of 105% were obtained. [ABSTRACT FROM AUTHOR]

An approach to improve the analytical performance of a Prussian Blue (PB)-based hydrogen peroxide transducer is described. In support of this objective, both the stabilizing and anti-interferent properties of nonconducting films were used. Electropolymerization on the top surface of PB modified electrodes is possible due to the high oxidizing ability of Berlin Green, and the growth of nonconductive polymers may be independently monitored by investigating the redox activity of the inorganic polycrystal. The best performance characteristics, which are advantageous over existing H2O2 sensors, were obtained for PB electrodes covered with electropolymerized o-phenylenediamine (1,2-diaminobenzene). The reported transducer remained at the 100% response state for more than 20 h under continuous flow of 0.1-mM hydrogen peroxide (flow rate 1 mlmin-1), which improves the stability level among the selective H2O2 sensors by one order of magnitude. The selectivity factor of the PB-poly (1,2-diaminobenzene) based transducer relative to ascorbate is nominally 600. PB-poly(1,2-diaminobenzene) modified electrode allows the detection hydrogen peroxide in the flow-injection mode down to 10-7 M with sensitivity of 0.3 AM-1cm-2, which is two times lower compared to the uncovered PB-based transducer. [ABSTRACT FROM PUBLISHER]

Describes methods for the construction of Prussian Blued-based carbon paste electrodes. Deposition of prussian blue on graphite powder; Electrodeposition of prussian blue on carbon paste; Modification of solid carbon paste electrodes by enzymes; Sensitivity and stability over wide range of hydrogen-ion concentration.

A facile “one-step” route to produce a homogenous and highly stable cobalt phthalocyanine (CoPc)-based dispersion by using carbon black (CB) as supporting material is reported. Herein, CB is proposed as effective material to load CoPc in order to obtain a CB/CoPc hybrid nanocomposite dispersion suitable for modifying screen-printed electrodes (SPEs) by an easy and automatable drop casting approach. CoPc resulted anchored to CB by a non-covalent physisorption, confirmed by IR and UV-visible spectroscopies, allowing to preserve the electrochemical performances of CoPc. The resulting CB/CoPc-modified SPE was tested as sensing tool to detect thiocholine, an enzymatic product of butyrylcholinesterase (BChE). The use of CB/CoPc leads to a highly sensitive thiocholine detection by applying a low potential (+0.05 V vs. internal reference) without fouling problem, a typical drawback that affects the thiol electrochemical detection. The favorable characteristics of the sensor were exploited for an easy BChE biosensor fabrication that renders this biosensor well suitable for mass-production. This electrochemical monoenzymatic biosensor was then challenged towards paraoxon, chosen as model organophosphorous pesticide, obtaining a low detection limit (18 nM). The suitability of the biosensor was tested in a waste water sample obtaining satisfactory recovery values, thus demonstrating its capability in such complex matrix. [ABSTRACT FROM AUTHOR]

A screen-printed electrode (SPE) modified with a carbon black (CB)-Au nanoparticles (AuNPs) composite is assembled and tested. Electrochemical and morphological investigations highlight the physico-chemical properties of the resulting AuNP-CB-SPE amperometric device with respect to SPEs modified with a single component of the nanocomposite. The effective performance of such a modified electrode in activating electrocatalytic processes, consisting both in oxidation and reduction reactions, is demonstrated. In particular, electrochemical tests on analytes such as glucose, hydrogen peroxide, hydroquinone, and ascorbic acid, evidence that the composite possesses electrocatalytic performance well superior with respect to the relevant mono-component modified SPE. As a consequence, a meaningful lowering of the peak potentials and improvement of the sensor sensitivities is observed when using AuNP-CB-SPEs with respect to both CB-SPEs and AuNP-SPEs. In the case of H 2 O 2 reduction, the occurrence of the electrochemical process at less negative potentials is coupled to an improvement of sensor sensitivity of about one order of magnitude. Concurrently, lower limit of detections, ranging from 20 to 99% less, have been obtained for the major part of the analytes studied, i.e. glucose, hydrogen peroxide and hydroquinone. Preliminary results reported here indicate that AuNP-CB-SPE can be proposed as an efficient amperometric sensor to be used in many analytical applications. [ABSTRACT FROM AUTHOR]

Highlights: [•] A novel laccase-modified screen printed electrode (SPE) has been developed. [•] Bionanocomposite sensor is optimized in terms of enzyme loading, pH and applied potential. [•] Linear range, sensitivity and LOD are assessed for BPA determination. [•] The biosensor has been challenged in tomato juice samples. [ABSTRACT FROM AUTHOR]

Abstract: A thionine-modified carbon paste electrode for catechol and Bisphenol A (BPA) detection is presented. Graphite powder was modified by adsorbing thionine as electrochemical mediator. The electrochemical response of the modified carbon paste electrode (CPE) was determined before electrode modification with tyrosinase. Then, tyrosinase was added in order to assemble a biosensor. Once established the best operative conditions, an interelectrode reproducibility around 7% was obtained and the resulting biosensor showed improved sensitivities and (S =139.6±1.1nA/μM for catechol and S =85.4±1.5nA/μM for BPA) in comparison with the biosensor constructed without thionine (S =104.4±0.5nA/μM for catechol and S =51.1±0.6nA/μM for BPA) and low detection limits (0.15μM for both the electrodes and analytes). Also the comparison with the results reported in the literature showed higher sensitivity and lower detection limit for our biosensor. Moreover the functioning of the thionine-tyrosinase CPE was validated following a biodegradation process of water polluted by BPA and comparing the time changes of BPA concentration inferred by the biosensor calibration curve and those determined by means of HPLC measurements. [Copyright &y& Elsevier]

Abstract: The aim of the present work was the development of an ELIME-array to achieve simple and rapid detection of AFB1 in corn samples. The system is based on an indirect competitive ELISA format using magnetic beads as immobilisation support and eight magnetised screen-printed electrodes as electrochemical transducers. After an optimisation study, a corn sample treatment, employing an extraction in acetonitrile/water followed by a clean-up step and solvent evaporation, was selected. For the construction of the calibration curve, which was used to evaluate both evaluation of the matrix effect on the performances of the ELIME-array and for the analysis of Certified Reference Materials (CRMs), standard solutions of AFB1 were added to blank dried corn extracts reconstituted in PBS. The detection limit and the sensitivity of the assay were calculated to be 0.6ngmL−1 and 1.5ngmL−1, respectively. Precision (11–26%) and recovery (95–114%) data of the ELIME-array, determined by analysing four CRMs, have shown that the proposed system appears suitable as a screening tool for the analysis of AFB1 in corn samples. [Copyright &y& Elsevier]