Electrochemical immunosensor for multiplexed detection of food-borne pathogens using nanocrystal bioconjugates and MWCNT screen-printed electrode

Bacterial food poisoning is an ever-present threat that can be prevented with proper care and handling of food products. A disposable electrochemical immunosensor for the simultaneous measurements of common food pathogenic bacteria namely Escherichia coli O157:H7 (E. coli), campylobacter and salmonella were developed. The immunosensor was fabricated by immobilizing the mixture of anti-E. coli, anticampylobacter and anti-salmonella antibodies with a ratio of 1:1:1 on the surface of the multiwall carbon nanotube-polyallylamine modified screen printed electrode (MWCNT-PAH/SPE). Bacteria suspension became attached to the immobilized antibodies when the immunosensor was incubated in liquid samples. The sandwich immunoassay was performed with three antibodies conjugated with specific nanocrystal ( -E. coli-CdS, -campylobacter-PbS and -salmonella-CuS) which has releasable metal ions for electrochemical measurements. The square wave anodic stripping voltammetry (SWASV) was employed to measure released metal ions from bound antibody nanocrystal conjugates. The calibration curves for three selected bacteria were found in the range of 1 × 103 – 5 × 105 cells mL−1 with the limit of detection (LOD) 400 cells mL−1 for salmonella, 400 cells mL−1 for campylobacter and 800 cells mL−1 for E. coli. The precision and sensitivity of this method show the feasibility of multiplexed determination of bacteria in milk samples.

Analysing organochlorine pesticides in strawberry jams using GC-ECD, GC-MS/MS and QuEChERS sample preparation

This paper describes a comparison of adaptations of the QuEChERS (quick, easy, cheap, effective, rugged and safe) approach for the determination of 14 organochlorine pesticide (OCP) residues in strawberry jam by concurrent use of gas chromatography (GC) coupled to electron capture detector (ECD) and GC tandem mass spectrometry (GC-MS/MS). Three versions were tested based on the original QuEChERS method. The results were good (overall average of 89% recoveries with 15% RSD) using the ultrasonic bath at five spiked levels. Performance characteristics, such as accuracy, precision, linear range, limits of detection (LOD) and quantification (LOQ), were determined for each pesticide. LOD ranged from 0.8 to 8.9 microg kg-1 ; LOQ was in the range of 2.5–29.8 microg kg- 1; and calibration curves were linear (r2>0.9970) in the whole range of the explored concentrations (5–100 microg kg- 1). The LODs of these pesticides were much lower than the maximum residue levels (MRLs) allowed in Europe for strawberries. The method was successfully applied to the quantification of OCP in commercially available jams. The OCPs were detected lower than the LOD.

Direct electroanalytical determination of fluvastatin in a pharmaceutical dosage form: batch and flow analysis

The reduction of luvastatin (FLV) at a hanging mercury-drop electrode (HMDE) was studied by square-wave adsorptive-stripping voltammetry (SWAdSV). FLV can be accumulated and reduced at the electrode, with a maximum peak current intensity at a potential of approximately 1.26V vs. AgCl=Ag, in an aqueous electrolyte solution of pH 5.25. The method shows linearity between peak current intensity and FLV concentration between 1.0 10 8 and 2.7 10 6 mol L 1. Limits of detection (LOD) and quantification (LOQ) were found to be 9.9 10 9 mol L 1 and 3.3 10 8 mol L 1, respectively. Furthermore, FLV oxidation at a glassy carbon electrode surface was used for its hydrodynamic monitoring by amperometric detection in a flow-injection system. The amperometric signal was linear with FLV concentration over the range 1.0 10 6 to 1.0 10 5 mol L 1, with an LOD of 2.4 10 7 mol L 1 and an LOQ of 8.0 10 7 mol L 1. A sample rate of 50 injections per hour was achieved. Both methods were validated and showed to be precise and accurate, being satisfactorily applied to the determination of FLV in a commercial pharmaceutical.

Electrochemical immunosensor for the analysis of the breast cancer biomarker HER2 ECD

Human epidermal growth factor receptor 2 (HER2) is a breast cancer biomarker that plays a major role in promoting breast cancer cell proliferation and malignant growth. The extracellular domain (ECD) of HER2 can be shed into the blood stream and its concentration is measurable in the serum fraction of blood. In this work an electrochemical immunosensor for the analysis of HER2 ECD in human serum samples was developed. To achieve this goal a screen-printed carbon electrode, modified with gold nanoparticles, was used as transducer surface. A sandwich immunoassay, using two monoclonal antibodies, was employed and the detection of the antibody–antigen interaction was performed through the analysis of an enzymatic reaction product by linear sweep voltammetry. Using the optimized experimental conditions the calibration curve (ip vs. log[HER2 ECD]) was established between 15 and 100 ng/mL and a limit of detection (LOD) of 4.4 ng/mL was achieved. These results indicate that the developed immunosensor could be a promising tool in breast cancer diagnostics, patient follow-up and monitoring of metastatic breast cancer since it allows quantification in a useful concentration range and has an LOD below the established cut-off value (15 ng/mL).

Sarcosine oxidase composite screen-printed electrode for sarcosine determination in biological samples

As the prostate cancer (PCa) progresses, sarcosine levels increase both in tumor cells and urine samples, suggesting that this metabolite measurements can help in the creation of non-invasive diagnostic methods for this disease. In this work, a biosensor device was developed for the quantification of sarcosine via electrochemical detection of H2O2 (at 0.6 V) generated from the catalyzed oxidation of sarcosine. The detection was carried out after the modification of carbon screen printed electrodes (SPEs) by immobilization of sarcosine oxidase (SOX) on the electrode surface. The strategies used herein included the activation of the carbon films by an electrochemical step and the formation of an NHS/EDAC layer to bond the enzyme to the electrode, the use of metallic or semiconductor nanoparticles layer previously or during the enzyme immobilization. In order to improve the sensor stability and selectivity a polymeric layer with extra enzyme content was further added. The proposed methodology for the detection of sarcosine allowed obtaining a limit of detection (LOD) of 16 nM, using a linear concentration range between 10 and 100 nM. The biosensor was successfully applied to the analysis of sarcosine in urine samples.

Sarcosine oxidase composite screen-printed electrode for sarcosine determination in biological samples

XIX Meeting of the Portuguese Electrochemical Society - XVI Iberic Meeting of Electrochemistry

Análise da interoperabilidade aplicada ao projeto BIM de Estruturas Metálicas

O BIM – Building Information Modeling – é um conceito de controlo e gestão de informação desenvolvida, entre diferentes especialidades e intervenientes envolventes, durante o ciclo de vida das construções. A utilização das tecnologias BIM, no ramo da Engenharia Mecânica e Civil, tem sido uma aposta constante, e cada vez mais concisa, nos projetos de construção. A justificação para a adoção destas metodologias mais eficientes, em substituição dos processos convencionais, prende-se com o facto desses processos convencionais, ainda hoje, apresentarem muitas dificuldades e problemas associados, por exemplo, à falta de comunicação entre os intervenientes e ao ineficiente controlo na gestão de projetos. O objetivo desta dissertação centra-se na análise da interoperabilidade de softwares BIM, ou seja, na verificação da viabilidade de exportação de dados dos modelos produzidos, entre as ferramentas BIM. Para este campo de ação contribuirá a análise da passagem de informação, relativa ao modelo de uma Nave Industrial modelada, em alguns dos softwares BIM, correntemente mais utilizados. O conhecimento adquirido com a modelação do caso de estudo do presente trabalho irá permitir identificar algumas lacunas existentes ao nível da falta de recomendações práticas que sirvam de orientação na modelação recorrendo a ferramentas informáticas BIM-compatíveis. Assim, a base deste trabalho consiste na criação de uma série de recomendações ou de um roteiro de modelação em específico para a disciplina de estruturas enquanto área de aplicação concreta do estudo efetuado. Nesse sentido, a proposta deste trabalho é de apresentar alguns critérios de modelação onde são definidos os elementos a modelar em cada fase do projeto.

POSTDATA – Towards publishing European poetry as linked open data

POSTDATA is a 5 year's European Research Council (ERC) Starting Grant Project that started in May 2016 and is hosted by the Universidad Nacional de Educación a Distancia (UNED), Madrid, Spain. The context of the project is the corpora of European Poetry (EP), with a special focus on poetic materials from different languages and literary traditions. POSTDATA aims to offer a standardized model in the philological field and a metadata application profile (MAP) for EP in order to build a common classification of all these poetic materials. The information of Spanish, Italian and French repertoires will be published in the Linked Open Data (LOD) ecosystem. Later we expect to extend the model to include additional corpora. There are a number of Web Based Information Systems in Europe with repertoires of poems available to human consumption but not in an appropriate condition to be accessible and reusable by the Semantic Web. These systems are not interoperable; they are in fact locked in their databases and proprietary software, not suitable to be linked in the Semantic Web. A way to make this data interoperable is to develop a MAP in order to be able to publish this data available in the LOD ecosystem, and also to publish new data that will be created and modeled based on this MAP. To create a common data model for EP is not simple since the existent data models are based on conceptualizations and terminology belonging to their own poetical traditions and each tradition has developed an idiosyncratic analytical terminology in a different and independent way for years. The result of this uncoordinated evolution is a set of varied terminologies to explain analogous metrical phenomena through the different poetic systems whose correspondences have been hardly studied – see examples in González-Blanco & Rodríguez (2014a and b). This work has to be done by domain experts before the modeling actually starts. On the other hand, the development of a MAP is a complex task though it is imperative to follow a method for this development. The last years Curado Malta & Baptista (2012, 2013a, 2013b) have been studying the development of MAP's in a Design Science Research (DSR) methodological process in order to define a method for the development of MAPs (see Curado Malta (2014)). The output of this DSR process was a first version of a method for the development of Metadata Application Profiles (Me4MAP) (paper to be published). The DSR process is now in the validation phase of the Relevance Cycle to validate Me4MAP. The development of this MAP for poetry will follow the guidelines of Me4MAP and this development will be used to do the validation of Me4MAP. The final goal of the POSTDATA project is: i) to be able to publish all the data locked in the WIS, in LOD, where any agent interested will be able to build applications over the data in order to serve final users; ii) to build a Web platform where: a) researchers, students and other final users interested in EP will be able to access poems (and their analyses) of all databases; b) researchers, students and other final users will be able to upload poems, the digitalized images of manuscripts, and fill in the information concerning the analysis of the poem, collaboratively contributing to a LOD dataset of poetry.