Fabiana Arduini

A thionine-modified carbon paste amperometric biosensor for catechol and bisphenol A determination.

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+/-1.1 nA/microM for catechol and S=85.4+/-1.5 nA/microM for BPA) in comparison with the biosensor constructed without thionine (S=104.4+/-0.5 nA/microM for catechol and S=51.1+/-0.6 nA/microM for BPA) and low detection limits (0.15 microM 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.

Recent advances in biosensors based on enzyme inhibition

Enzyme inhibitors like drugs and pollutants are closely correlated to human and environmental health, thus their monitoring is of paramount importance in analytical chemistry. Enzymatic biosensors represent cost-effective, miniaturized and easy to use devices; particularly biosensors based on enzyme inhibition are useful analytical tools for fast screening and monitoring of inhibitors. The present review will highlight the research carried out in the last 9 years (2006-2014) on biosensors based on enzyme inhibition. We underpin the recent advances focused on the investigation in new theoretical approachs and in the evaluation of biosensor performances for reversible and irreversible inhibitors. The use of nanomaterials and microfluidic systems as well as the applications of the various biosensors in real samples is critically reviewed, demonstrating that such biosensors allow the development of useful devices for a fast and reliable alarm system.

Development of a bio-electrochemical assay for AFB1 detection in olive oil.

A novel biosensor assay format for aflatoxin based on acetylcholinesterase (AChE) inhibition by aflatoxin B(1) (AFB(1)) is proposed. The AChE was present in solution and an amperometric choline oxidase biosensor was used for monitoring its residual activity. To create the biosensor, the choline oxidase was immobilized by cross-linking onto screen-printed electrodes modified with Prussian Blue (PB) and these were used to detect the H(2)O(2) at low potential (-0.05V versus a screen-printed internal silver pseudoreference electrode). For the development of the AFB(1) assay, several parameters such as AChE and substrate concentration, the methanol effect, and pH were evaluated and optimized. The linear working range was assessed to be 10-60ppb. Concentrations as low as 2ppb, which correspond to the legal limit of AFB(1) in food for humans, were detected after a pre-concentration step. The suitability of the method was evaluated using commercial olive oil samples. A recovery equal to 78+/-9% for 10ppb of AFB(1) in olive oil samples was obtained.

Nanomaterials in electrochemical biosensors for pesticide detection: advances and challenges in food analysis

AbstractThis overview (with 114 refs.) covers the progress made between 2010 and 2015 in the field of nanomaterial based electrochemical biosensors for pesticides in food. Its main focus is on strategies to analyze real samples. The review first gives a short introduction into the most often used biorecognition elements. These include (a) enzymes (resulting in inhibition-based and direct catalytic biosensors), (b) antibodies (resulting in immunosensors), and (c) aptamers (resulting in aptasensors). The next main section covers the various kinds of nanomaterials for use in biosensors and includes carbonaceous species (carbon nanotubes, graphene, carbon black and others), and non-carbonaceous species in the form of nanoparticles, rods, or porous materials. Aspects of sample treatment and real sample analysis are treated next before discussing vanguard technologies in tailor-made food analysis. Graphical abstractLast trends made between 2010 and 2015 on the use of nanomaterials, including graphene, carbon nanotubes, carbon black, metallic nanoparticles, for the development of enzymatic biosensors, immunosensors, and aptasensors were reported, tackling the issues related to pesticide detection in agrifood sector.

Reversible Enzyme Inhibition–Based Biosensors: Applications and Analytical Improvement Through Diagnostic Inhibition

Abstract The present review reports the research carried out during past 9 years on biosensors based on reversible enzyme inhibition for determination of drugs, pollutants, and toxic compounds. Applications in food, environmental, and pharmaceutical fields are also reported. Special attention is paid to the optimization of parameters such as enzyme immobilization, substrate concentration, and incubation time. On the basis of the studies reviewed here, it is our view that enzyme inhibition–based biosensors have been shown to be useful analytical tools.

Screen-printed electrode modified with carbon black nanoparticles for phosphate detection by measuring the electroactive phosphomolybdate complex

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.

Electroanalytical Characterization of Carbon Black Nanomaterial Paste Electrode: Development of Highly Sensitive Tyrosinase Biosensor for Catechol Detection

In this work, the electrochemical behavior of carbon black paste electrode prepared using a nanostructured commercial carbon black (N220) was investigated. The sensor was challenged with several potentially interesting analytes by means of cyclic voltammetry technique and the results compared with graphite carbon paste electrode. Shifting in peak potential and/or increase in the peak currents for some analytes such as ferricyanide, ascorbic acid, acetoaminophen, epinephrine, and DOPAC were observed. The carbon black paste was combined with tyrosinase enzyme to produce a biosensor which was challenged in amperometric mode with catechol. The highest sensitivity, equal to 625 nA/μM, coupled with lowest detection limit of 0.008 μM was observed for this formulation relative to those made with graphite and even when compared with carbon nanotubes tyrosinase paste electrode previously reported. In this way, the carbon black could be considered a good electrode material for constructing other electrochemical biosensors with the advantage to be a nanostrutured material at low cost.

Extraction and Detection of Pesticides by Cholinesterase Inhibition in a Two‐Phase System: a Strategy to Avoid Heavy Metal Interference

Abstract A novel method of extraction has been developed to avoid the presence of heavy metals during the measurement of pesticides based on acetylcholinesterase (AchE) inhibition. Heavy metals have been in fact demonstrated in this article to interfere when the assay is performed by using the classic spectrophotometric Ellmans method. We present the results obtained with an assay system using two different phases, one organic and the other aqueous, in which the pesticide and the enzyme are, respectively, solubilized. In a first step, the concentration of the substrate acetylthiocholine (1 mM), of the enzyme (7 mU mL−1), and the reaction time (20 min) for measurement of enzyme activity were optimized in aqueous solution. Next, the effect of an organic phase on the enzyme activity was studied by the addition of various solvents with the activity being evaluated after 10 min of mixing. It was found that by using hexane, the enzyme retained almost 100% of its activity, and this solvent was chosen for further development of the pesticide assay. Hexane was spiked with different concentrations of pesticides and then added to the enzyme aqueous phase. The pesticides were shown to be able to inhibit the enzyme by interaction at the interface between the two solutions. The degree of inhibition obtained with increasing amounts of pesticide was evaluated. A 50% inhibition was observed for a paraoxon solution of 9×10−7 M.

Development of a Hydrogen Peroxide Sensor Based on Screen-Printed Electrodes Modified with Inkjet-Printed Prussian Blue Nanoparticles

A sensor for the simple and sensitive measurement of hydrogen peroxide has been developed which is based on screen printed electrodes (SPEs) modified with Prussian blue nanoparticles (PBNPs) deposited using piezoelectric inkjet printing. PBNP-modified SPEs were characterized using physical and electrochemical techniques to optimize the PBNP layer thickness and electroanalytical conditions for optimum measurement of hydrogen peroxide. Sensor optimization resulted in a limit of detection of 2 × 10−7 M, a linear range from 0 to 4.5 mM and a sensitivity of 762 μA·mM−1·cm−2 which was achieved using 20 layers of printed PBNPs. Sensors also demonstrated excellent reproducibility (<5% rsd).

Fully integrated ready-to-use paper-based electrochemical biosensor to detect nerve agents

Paper-based microfluidic devices are gaining large popularity because of their uncontested advantages of simplicity, cost-effectiveness, limited necessity of laboratory infrastructure and skilled personnel. Moreover, these devices require only small volumes of reagents and samples, provide rapid analysis, and are portable and disposable. Their combination with electrochemical detection offers additional benefits of high sensitivity, selectivity, simplicity of instrumentation, portability, and low cost of the total system. Herein, we present the first example of an integrated paper-based screen-printed electrochemical biosensor device able to quantify nerve agents. The principle of this approach is based on dual electrochemical measurements, in parallel, of butyrylcholinesterase (BChE) enzyme activity towards butyrylthiocholine with and without exposure to contaminated samples. The sensitivity of this device is largely improved using a carbon black/Prussian Blue nanocomposite as a working electrode modifier. The proposed device allows an entirely reagent-free analysis. A strip of a nitrocellulose membrane, that contains the substrate, is integrated with a paper-based test area that holds a screen-printed electrode and BChE. Paraoxon, chosen as nerve agent simulant, is linearly detected down to 3µg/L. The use of extremely affordable manufacturing techniques provides a rapid, sensitive, reproducible, and inexpensive tool for in situ assessment of nerve agent contamination. This represents a powerful approach for use by non-specialists, that can be easily broadened to other (bio)systems.