Sandra A. V. Eremia

Disposable biosensor based on platinum nanoparticles-reduced graphene oxide-laccase biocomposite for the determination of total polyphenolic content

A disposable amperometric biosensor was developed for the detection of total polyphenolic compounds from tea infusions. The biosensor was designed by modifying the surface of a carbon screen-printed electrode with platinum nanoparticles and reduced graphene oxide, followed by the laccase drop-casting and stabilization in neutralised 1% Nafion solution. The obtained biosensor was investigated by scanning electron microscopy and electrochemical techniques. It was observed that platinum nanoparticles-reduced graphene oxide composite had synergistic effects on the electron transfer and increased the electroactive surface area of the carbon screen-printed electrode. The constructed analytical tool showed a good linearity in the range 0.2-2 μM for caffeic acid and a limit of detection of 0.09 μM. The value of Michaelis-Menten apparent constant was calculated from the electrochemical version of Lineweaver-Burk equation to be 2.75 μM. This disposable laccase biosensor could be a valuable tool for the estimation of total polyphenolic content from tea infusions.

Molybdenum disulphide and graphene quantum dots as electrode modifiers for laccase biosensor

A nanocomposite formed from molybdenum disulphide (MoS2) and graphene quantum dots (GQDs) was proposed as a novel and suitable support for enzyme immobilisation displaying interesting electrochemical properties. The conductivity of the carbon based screen-printed electrodes was highly improved after modification with MoS2 nanoflakes and GQDs, the nanocomposite also providing compatible matrix for laccase immobilisation. The influence of different modification steps on the final electroanalytical performances of the modified electrode were evaluated by UV-vis absorption and fluorescence spectroscopy, scanning electron microscopy, transmission electron microscopy, X ray diffraction, electrochemical impedance spectroscopy and cyclic voltammetry. The developed laccase biosensor has responded efficiently to caffeic acid over a concentration range of 0.38-100µM, had a detection limit of 0.32µM and a sensitivity of 17.92nAµM(-1). The proposed analytical tool was successfully applied for the determination of total polyphenolic content from red wine samples.

Laccase-Nafion based biosensor for the determination of polyphenolic secondary metabolites.

A laccase-based biosensor was developed by specific enzyme adsorption on screen-printed working electrodes of DROPSENS cells, and stabilized with Nafion 0.1% membrane. The electrode was characterized with respect to response time, sensitivity, linear range, detection limit, pH dependence, interferences, and long-term stability. The tested substrates were catechol, rosmarinic acid, caffeic acid, chlorogenic acid, and gallic acid. The optimized biosensor proved the following characteristic performances: the apparent Michaelis Menten calculated considering rosmarinic acid substrate 8.3 × 10−6 mol L−1 (r = 0.995, n = 6); the dynamic range of biosensor response for rosmarinic acid 7 × 10−7 − 1.5 × 10−6 mol L−1; the detection limit for rosmarinic acid 1.19 × 10−7 mol L−1 (RSD = 1.08%, n = 3). It was noticed that the biosensor reaches systematically 90% to 94.3% from the response obtained by LC-DAD-ESI-MS for real samples.

Disposable dual sensor array for simultaneous determination of chlorogenic acid and caffeine from coffee

In this work a novel sensor array platform based on a dual carbon screen-printed electrode was developed for the simultaneous determination of chlorogenic acid and caffeine. One of the carbon working electrodes was modified with platinum nanoparticles, reduced graphene oxide and laccase (C-SPE/Pt-NPs/RGO/lacc-biosensor) for chlorogenic acid determination and the second carbon working electrodes was modified with reduced graphene oxide and Nafion (C-SPE/RGO/Nafion-sensor) for caffeine determination. Cyclic voltammetry was used to characterise and optimise the dual sensor array while chronoamperometry was used to investigate the bioelectrocatalytic response. The C-SPE/Pt-NPs/RGO/lacc for biosensing chlorogenic acid exhibited a sensitivity of 0.02 μA μM−1 and a detection limit of 2.67 μM whereas the C-SPE/RGO/Nafion used for sensing caffeine has showed a sensitivity of 1.38 μA μM−1 and a detection limit of 0.22 μM. The developed sensor array was used to determine these two major coffee bean compounds from real coffee samples. Due to its simplicity, feasibility and accessibility, the developed dual sensor array could represent the basis of a valuable analytical tool able to screen both chlorogenic acid and caffeine content from coffee samples offering important information about the phytochemical composition of the samples.

Biosensors for the Determination of Phenolic Metabolites

Antioxidants are groups of chemical substances, the most abundant being polyphenols, mainly found in plants, fruits and vegetables. They include flavonoids, flavonoid derivatives, polyphenols, carotenoids and anthocyanins. Currently, the nutritional quality of many foodstuffs is guaranteed by the presence of antioxidant compounds. The importance of these chemicals as indicators and preservatives of nutritional quality makes necessary the development of accurate, versatile and rapid analytical tools necessary to detect their presence in many foodstuffs and to assess their antioxidant efficacy. In this chapter, enzyme-based biosensors such as monophenol monooxygenase (tyrosinase), catechol oxidase (laccase) and horseradish peroxidase (HRP) are reviewed. Actually, these biosensors are the most commonly used for the detection of polyphenols and flavonoids content.

Probiotic Strains Influence on Infant Microbiota in the In Vitro Colonic Fermentation Model GIS1.

AbstractThe main goal of our study was to evaluate the effect of the individual administration of five lyophilized lactic acid bacteria strains (Lactobacillus fermentum 428ST, Lactobacillus rhamnosus E4.2, Lactobacillus plantarum FCA3, Lactobacillus sp. 34.1, Weissella paramesenteroides FT1a) against the in vitro simulated microbiota of the human colon using the GIS1 system. The influence on the metabolic activity was also assessed by quantitative determination of proteins and polysaccharides at each segment of human colon. The obtained results indicated that the lactic acid bacteria L. rhamnosus E4.2 and W. paramesenteroides FTa1 had better efficiency in synthesising exopolysaccharides and also a better probiotic potential and therefore could be recommended for use in probiotics products or food industry.

Development of a nanocomposite system and its application in biosensors construction

AbstractThe present work describes the development of a nanocomposite system and its application in construction of a new amperometric biosensor applied in the determination of total polyphenolic content from propolis extracts. The nanocomposite system was based on covalent immobilization of laccase on functionalized indium tin oxide nanoparticles and it was morphologically and structural characterized. The casting of the developed nanocomposite system on the surface of a screen-printed electrode was used for biosensor fabrication. The analytical performance characteristics of the settled biosensor were determined for rosmarinic acid, caffeic acid and catechol (as laccase specific substrate). The linearity was obtained in the range of 1.06×10−6 − 1.50×10−5 mol L−1 for rosmarinic acid, 1.90×10−7 − 2.80×10−6 mol L−1 for caffeic acid and 1.66×10−6 − 7.00×10−6 mol L−1 for catechol. A good sensitivity of amperometric biosensor 141.15 nA µmol−1 L−1 and fair detection limit 7.08×10−8 mol L−1 were obtained for caffeic acid. The results obtained for polyphenolic content of propolis extracts were compared with the chromatographic data obtained by liquid-chromatography with diode array detection.

Monitoring of Rosmarinic Acid Accumulation in Sage Cell Cultures using Laccase Biosensor

INTRODUCTION A recently developed laccase based biosensor is used for polyphenols determination from in vitro Salvia cultures, the results being expressed as rosmarinic acid equivalent content. OBJECTIVE The aim of this work was to use a previously developed laccase biosensor for the determination of total phenolic content from in vitro cultivated Salvia, and to support the biosensors further application for the assessment of polyphenols metabolites. METHODOLOGY The biosensor was constructed by drop casting 3 μL of laccase solution and stabilisation with 0.1 % Nafion solution onto a DropSens carbon screen-printed electrode. Electrochemical measurements were carried out in a 0.1 mol/L phosphate buffer (pH 4.50), the applied working potential being -30 mV versus reference electrode. RESULTS The response of the biosensor developed was characterised in terms of repeatability, accuracy and precision; the limit of detection was 7.5 × 10(-7) mol/L, the limit of determination was 9.5 × 10⁻⁷ mol/L, and linear response range for rosmarinic acid was 1 × 10⁻⁶-10⁻⁵ mol/L. CONCLUSION A stable, sensitive and simple biosensor based on laccase-nafion was used for monitoring the total polyphenolic content from two in vitro cultivated plants. The biosensor response was free of electrochemical interferences and of possible interferences from growth media constituents, demonstrating a high sensitivity for rosmarinic acid determination in cell culture suspensions.

Phenolic and Anthocyanin Profile of Valea Calugareasca Red Wines by HPLC-PDA-MS and MALDI-TOF Analysis

The quality of red wines is given by phenolic compounds and anthocyanins and is associated with colour, taste and therapeutic effects on human health. This work aims to provide a detailed profile of phenolic compounds and anthocyanins that are found in five red wine samples from Dealu Mare-Valea Calugareasca region. The phenolic and anthocyanin profiles of the red wine samples were determined by high-performance liquid chromatography-photodiode array-mass spectrometry and matrix-assisted laser desorption/ionisation–time of flight, respectively. The results obtained showed that Feteasca Neagra had the highest content of phenolic compounds followed by Pinot Noir while the preponderant compound was gallic acid. Amongst anthocyanins, malvidin was found to be the major compound and the highest anthocyanin content was found also for Feteasca Neagra wine. A simple high-performance liquid chromatography-photodiode array-mass spectrometry method was developed, optimised and applied for the quantification of phenolic compounds in red wine samples from Dealu Mare-Valea Calugareasca region. In the same time, a rapid matrix-assisted laser desorption/ionisation–time of flight method that does not need sample preparation was applied for the identification of anthocyanins. Moreover, the phenolic and anthocyanin composition of red wines from Dealu Mare-Valea Calugareasca region is reported for the first time. The phenolic and anthocyanin profile determination will be beneficial for the Romanian winemakers to produce high-quality red wines.

Application of an optimized electrochemical sensor for monitoring astaxanthin antioxidant properties against lipoperoxidation

An optimized electrochemical sensor was developed to assess the antioxidant capacity of carotenoids, accumulating during the life cycle of Haematococcus pluvialis cell cultures. The sensor was improved with a composite renewable surface made of immobilised phosphatidylcholine (PC) on magnetic nanobeads of iron oxide (Fe3O4) and PC/Fe3O4, and it was used to monitor the antioxidant properties of the ketocarotenoid astaxanthin against in situ generated phosphatidylcholine lipoperoxides. The surface configuration was able to mimic the natural position and orientation of astaxanthin in the cellular membrane, conferring to the whole experimental set-up good sensitivity for reactive oxygen species (limit of detection for peroxyl radicals 9.1 × 10−10 mol L−1) with a linear response ranging between 10−8 and 1.6 × 10−6 mol L−1. The sensor has been proved suitable for both batch and flow measurements. The accuracy of the flow measurements was unaffected by the magnetic field intensity. Electrochemical measurements confirmed that natural astaxanthin is a more effective antioxidant than synthetic astaxanthin, vitamin E and lutein and the protective effect of astaxanthin correlates with its concentration inside the cell. The newly developed sensor is hence useable for in-line monitoring of whole-cell based industrial bioprocesses for the production of astaxanthin.