Bioelectronic tongue based on lipidic nanostructured layers containing phenol oxidases and lutetium bisphthalocyanine for the analysis of grapes

Producción CientíficaIn this work, a multisensor system formed by nanostructured voltammetric biosensors based on phenol oxidases (tyrosinase and laccase) has been developed. The enzymes have been incorporated into a biomimetic environment provided by a Langmuir-Blodgett (LB) film of arachidic acid (AA). Lutetium bisphthalocyanine (LuPc2) has also been introduced in the films to act as electron mediator. The incorporation of the enzymes to the floating layers to form Tyr/AA/LuPc2 and Lac/AA/LuPc2 films has been confirmed by the expansion in the surface pressure isotherms and by the AFM images. The voltammetric response towards six phenolic compounds demonstrates the enhanced performance of the biosensors that resulted from a preserved activity of the tyrosinase and laccase combined with the electron transfer activity of LuPc2. Biosensors show improved detection limits in the range of 10-7-10-8molL-1. An array formed by three sensors AA/LuPc2, Tyr/AA/LuPc2 and Lac/AA/LuPc2 has been employed to discriminate phenolic antioxidants of interest in the food industry. The Principal Component Analysis scores plot has demonstrated that the multisensor system is able to discriminate phenols according to the number of phenolic groups attached to the structure. The system has also been able to discriminate grapes of different varieties according to their phenolic content. This good performance is due to the combination of four factors: the high functionality of the enzyme obtained using a biomimetic immobilization, the signal enhancement caused by the LuPc2 mediator, the improvement in the selectivity induced by the enzymes and the complementary activity of the enzymatic sensors demonstrated in the loading plots.Ministerio de Economía, Industria y Competitividad – FEDER (Grant CICYT AGL2012-33535)Junta de Castilla y León (programa de apoyo a proyectos de investigación - Ref. VA-032U13

Array of biosensors for discrimination of grapes according to grape variety, vintage and ripeness.

A bioelectronic tongue based on nanostructured biosensors specific for the simultaneous detection of sugars and phenols has been developed. The array combined oxidases and dehydrogenases immobilized on a lipidic layer prepared using the Langmuir-Blodgett technique where Glucose oxidase, d-Fructose dehydrogenase, Tyrosinase or Laccase were imbibed. A phthalocyanine was co-immobilized in the sensing layer and used as electron mediator. The array thus formed has been used to analyze grapes and provides global information about the samples while providing specific information about their phenolic and their sugar content. Using Principal Component Analysis (PCA) the array of voltammetric biosensors has been successfully used to discriminate musts prepared from different varieties of grapes (Tempranillo, Garnacha, Cabernet-Sauvignon, Prieto Picudo and Mencía). Differences could be also detected between grapes of the same variety and cultivar harvested in two successive vintages (2012 and 2013).2020-07-092020-07-0

The advantages of disposable screen-printed biosensors in a bioelectronic tongue for the analysis of grapes

Producción CientíficaDisposable screen-printed sensors have been modified with enzymes and used to form a bioelectronic tongue dedicated to the discrimination between different grape varieties. The multisensory system combined serigraphied electrodes modified with carbon, platinum, gold, graphene, Prussian blue and nickel oxide nanoparticles (M-SPE) covered with glucose oxidase (M-GOX-SPE) or tyrosinase (M-Tyr-SPE). The M-GOX-SPE and M-Tyr-SPE sensors produced a variety of responses due to the different behavior of the electron mediators of the six screen-printed materials used for the electro-catalysis of the glucose and phenols by means of glucose oxidase and tyrosinase. This variety of responses, together with the capability of the sensors to detect glucose or phenols, allowed the bioelectronic tongue developed here to discriminate between the juices obtained from different varieties of grape. Partial least-squares (PLS-1) multivariate calibration of electrochemical data has been successfully applied to the simultaneous determination of glucose and polyphenols in musts. The discrimination capability shown by this array of cheap and single-use sensors was similar to that found in other complex bioelectronic tongues. The lower price, ease of use and portability of the modified screen-printed electrode system makes the bioelectronic tongue developed here an alternative tool that can be used in situ in the vineyard block.Ministerio de Economía, Industria y Competitividad – FEDER (Grant CICYT AGL2012-33535)Junta de Castilla y León (programa de apoyo a proyectos de investigación - Ref. VA-032U13)University of Valladolid (PIF-UVa

Electronic Noses and Tongues in Wine Industry

Producción CientíficaThe quality of wines is usually evaluated by a sensory panel formed of trained experts or traditional chemical analysis. Over the last few decades, electronic noses (e-noses) and electronic tongues have been developed to determine the quality of foods and beverages. They consist of arrays of sensors with cross-sensitivity, combined with pattern recognition software, which provide a fingerprint of the samples that can be used to discriminate or classify the samples. This holistic approach is inspired by the method used in mammals to recognize food through their senses. They have been widely applied to the analysis of wines, including quality control, aging control, or the detection of fraudulence, among others. In this paper, the current status of research and development in the field of e-noses and tongues applied to the analysis of wines is reviewed. Their potential applications in the wine industry are described. The review ends with a final comment about expected future developments.CM-P agradece a la Universidad de Valladolid por su beca PIF-UVa y CG-H por su contrato pre-doctoral JCYL (BOCYL-D-24112015-9).Ministerio de Economía, Industria y Competitividad – FEDER (Grant AGL2015-67482-R)Junta de Castilla y León (programa de apoyo a proyectos de investigación - Ref. VA-032U13

An Electrochemical Quartz Crystal Microbalance Multisensor System Based on Phthalocyanine Nanostructured Films: Discrimination of Musts

Producción CientíficaAn array of electrochemical quartz crystal electrodes (EQCM) modified with nanostructured films based on phthalocyanines was developed and used to discriminate musts prepared from different varieties of grapes. Nanostructured films of iron, nickel and copper phthalocyanines were deposited on Pt/quartz crystals through the Layer by Layer technique by alternating layers of the corresponding phthalocyanine and poly-allylamine hydrochloride. Simultaneous electrochemical and mass measurements were used to study the mass changes accompanying the oxidation of electroactive species present in must samples obtained from six Spanish varieties of grapes (Juan García, Prieto Picudo, Mencía Regadío, Cabernet Sauvignon, Garnacha and Tempranillo). The mass and voltammetric outputs were processed using three-way models. Parallel Factor Analysis (PARAFAC) was successfully used to discriminate the must samples according to their variety. Multi-way partial least squares (N-PLS) evidenced the correlations existing between the voltammetric data and the polyphenolic content measured by chemical methods. Similarly, N-PLS showed a correlation between mass outputs and parameters related to the sugar content. These results demonstrated that electronic tongues based on arrays of EQCM sensors can offer advantages over arrays of mass or voltammetric sensors used separately.Ministerio de Economía, Industria y Competitividad – FEDER (Grant CICYT AGL2012-33535)Junta de Castilla y León (programa de apoyo a proyectos de investigación - Ref. VA-032U13)University of Valladolid (PIF-UVa

Improvement of electrocatalytic effect in voltammetric sensors based on phthalocyanines

Producción CientíficaVoltammetric sensors based on phthalocyanines have been used to detect a variety of compounds. In this paper, the state of the art of sensors prepared using classical techniques will be revised. Then, new strategies to improve the performance of the sensors will be described using as example sensors chemically modified with lutetium bisphthalocyanine (LuPc2) dedicated to the detection of phenols of interest in the food industry. Classical LuPc2 carbon paste electrodes can detect phenols such as catechol, caffeic acid or pyrogallol with limits of detection in the range of 10-4–10-5 M. The performance can be improved by using nanostructured Langmuir–Blodgett (LB) or Layer by Layer (LbL) films. The enhanced surface to volume ratio produce an increase in the sensitivity of the sensors. Limits of detection of 10-5–10-7 M are attained, which are one order of magnitude lower than those obtained using conventional carbon paste electrodes. Moreover, these techniques can be used to coimmobilize two electrocatalytic materials in the same device. The limits of detection obtained in LB sensors combining LuPc2/AuNPs or LuPc2/CNT are further improved. Finally, the LB technique has been used to prepare biosensors where a phenol oxydase (such as tyrosinase or lacasse) is immobilized in a biomimetic environment that preserves the enzymatic activity. Moreover, LuPc2 can be co-immobilized with the enzyme in a lipidc film formed by arachidic acid (AA). LuPc2 can act as an electron mediator facilitating the electron transfer. These biomimetic sensors formed by LuPc2/AA/enzyme show Limits of detection of 10-8 M and an enhanced selectivity.CM-P agradece a la Universidad de Valladolid por su beca PIF-UVa y CG-H por su contrato pre-doctoral JCYL (BOCYL-D-24112015-9).Ministerio de Economía, Industria y Competitividad – FEDER (Grant CICYT AGL2012-33535)Junta de Castilla y León (programa de apoyo a proyectos de investigación - Ref. VA-032U13

Mechanical Behaviour at Low Strains of LDPE Foams with Cell Sizes in the Microcellular Range: Advantages of Using these Materials in Structural Elements.

This paper presents the production method and the compressive mechanical response at low strains for a collection of polyethylene foams with high densities and cell sizes in the microcellular range. The materials were produced using a modified compression moulding technique that allows and independent control of density and cell size. The materials had a relative density between 0.27 and 0.92, an homogeneous and multi-structured cellular structure with dense skin and foamed core and cell sizes in the range 30 to 100 microns. The YoungŽs modulus was reduced when density did. For relative densities higher than 0.7, the reduced Young¿s modulus of the foams was higher than that of the solid. In addition, it has been proved that variations in the cell size at constant density did not influence the Young¿s modulus. The advantages of using these materials for the production of plastic pipes have been analysed. A reduction of the weight of pipes loaded in compression of up to 42 % can be reached by using these foams in spite of the solid material from which the foam was produced

Nanostructured thin films based on phthalocyanines: electrochromic displays and sensors

Producción CientíficaThe group of the University of Valladolid is a multidisciplinary team formed by chemists, physicists and engineers. The activities of the group are focused to the study of the physicochemical properties of nanostructured Langmuir-Blodgett thin films based on phthalocyanines and their applications. Films of a variety of phthalocyanine molecules including several metallophthalocyanines, lanthanide double decker phthalocyanines and heteroleptic derivatives have been prepared. Their spectroelectrochemical properties have been described in detail and compared with those observed in disordered casted films or microcrystalline evaporated films. The group has dedicated special attention to films based on rare earth double decker compounds due to their unique semiconducting, optical and electrochemical properties. A rich electrochromism has been demonstrated in thin films of this family of compounds. The reversibility is improved in nanostructured Langmuir-Blodgett films. This has permitted development of an electrochromic display that can change its color from blue to green and finally to red. At the present moment, our main objective is the design of sensors able to detect gases and liquids. It has been demonstrated that thin film assemblies based on rare earth bisphthalocyanines modify their conductivity and their optical properties in the presence of electron donor or electron acceptors gases. Changes are also observed when the devices are exposed to Volatile Organic Compounds such as esters, alcohols or aldehydes which are responsible of odors in foods and beverages. Liquid sensors have also been developed. Their working principle is based in the fact that the rich electrochemical properties of phthalocyanine thin films are extremely sensitive to the nature of the electrolytic solution. Arrays of phthalocyanines have been used to construct an electronic nose able to discriminate odors from a variety of foods and beverages. Similarly, phthalocyanines have also been used to construct an electronic tongue based on voltammetric sensors. This is one of the main contributions of the group to the field of sensors.2022-07-62022-07-0

Amperometric tyrosinase based biosensor using an electropolymerized phosphate-doped polypyrrole film as an immobilization support. Application for detection of phenolic compounds

Producción CientíficaAn amperometric biosensor was constructed by immobilization of enzyme, tyrosinase, in an electrochemically synthesized phosphate ion-doped polypyrrole film on a Pt disk electrode. The tyrosinase maintains its bioactivity well within the polypyrrole thin film. A clearly defined reduction current proportional to the phenolic compounds concentration was observed in cyclic voltammetry, which attributed to the reduction of enzymatically produced quinone at the electrode surface. Phenolic compounds were quantitatively estimated in aqueous medium by the direct electrochemical reduction of enzymatically liberated quinone species at −0.05 V by chronoamperometry. For the all six phenolic compounds analyzed, the kinetics of the enzymatic reaction fitted into a Michaelis–Menten type kinetics, as demonstrated by the h parameter close to 1 obtained from the Hill's plot. The sensitivity followed the decreasing order catechol > phenol > 2-bromophenol > 2-chlorophenol > 2-iodophenol > 2-fluorophenol. The greater value of Imax and the lowest was found for catechol. The detection limits were in the range of 0.84–8.54 μM. The lowest detection limits were found for catechol and the highest for 2-fluorophenol.2022-07-62022-07-0

Films of lutetium bisphthalocyanine nanowires as electrochemical sensors

Producción CientíficaLutetium bisphthalocyanine (LuPc(2)) nanowires have been successfully obtained by electrophoretic deposition (EPD). The influence of the deposition conditions and annealing in the structure of the films has been studied by AFM, SEM, X-ray diffraction (XRD), UV-vis absorption, and near-infrared (NIR). The electrochemical properties of the EDP films immersed in different electrolytic solutions (KCl, MgCl(2), KClO(4), HCl, and NaOH) indicate that anions diffuse inside the film to maintain the electroneutrality and the kinetics follows the Randles-Sevcik equation. The stability of the response increases strongly upon annealing due to the improvement of the adhesion of the sensitive material to the substrate. The EPD films have been successfully used to detect caffeic acid (an antioxidant of interest in the food industry). The anodic peak associated with the oxidation of caffeic acid appears at 0.54 V and is linearly dependent on the caffeic acid concentration in the 6 × 10(-5) M to 5 × 10(-4) M range with a detection limit of 3.12 × 10(-5) M. The electrochemical behavior of the annealed LuPc(2) EPD films is similar to that observed using Langmuir-Blodgett (LB) nanostructured films. However, the different molecular organization of the molecules inside the film causes differences in the shape and position of the peaks. Although LuPc(2) sensors prepared with both EPD and LB techniques provide stable and reproducible responses, the use of EPD is preferred for real sensing applications because of its lower cost, shorter preparation time, and longer lifetime.2022-07-62022-07-0