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1.
Sci Rep ; 10(1): 21077, 2020 12 03.
Artigo em Inglês | MEDLINE | ID: mdl-33273679

RESUMO

The tellurium doped zinc imidazole framework (Te@ZIF-8) is prepared by a two-step hydrothermal strategy for the electrochemical sensing of hydrogen peroxide. Material is characterized by transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). The electrochemical characterization of the MOF modified electrode is done by a three-electrode system. Electrochemical sensing of hydrogen peroxide is made by cyclic voltammetry, amperometry, and impedance measurements. Results demonstrate that Te@ZIF-8 shows a detection limit of 60 µM with linearity up to 0.98855. Material is stable to 1000 cycles with no significant change in electrochemical response. Amperometry depicts the recovery of hydrogen peroxide from human serum up to 101%. Impedance curve reveals the surface of Te@ZIF-8-GCE (glassy carbon electrode) as porous and rough and an interface is developed between analyte ions and the sensing material. Finally, the modified electrode is used for the quantitative determination of hydrogen peroxide from serum samples of pancreatic cancer patients, diagnosed with CA 19-9.


Assuntos
Técnicas Eletroquímicas/métodos , Peróxido de Hidrogênio/sangue , Estruturas Metalorgânicas/química , Neoplasias Pancreáticas/sangue , Telúrio/química , Zinco/química , Análise Química do Sangue/instrumentação , Análise Química do Sangue/métodos , Análise Química do Sangue/normas , Células Cultivadas , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/normas , Eletrodos , Humanos , Imidazóis/química , Limite de Detecção
2.
Biosens Bioelectron ; 155: 112111, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32217334

RESUMO

Electrochemical biosensors possess numerous desirable qualities for target detection, such as portability and ease of use, and are often considered for point-of-care (POC) development. Label-free affinity electrochemical biosensors constructed with semiconductor manufacturing technology (SMT)-produced electrodes and a streptavidin biomediator currently display the highest reproducibility, accuracy, and stability in modern biosensors. However, such biosensors still do not meet POC guidelines regarding these three characteristics. The purpose of this research was to resolve the limitations in reproducibility and accuracy caused by problems with production of the biosensors, with the aim of developing a platform capable of producing devices that exceed POC standards. SMT production settings were optimized and bioreceptor immobilization was improved through the use of a unique linker, producing a biosensor with exceptional reproducibility, impressive accuracy, and high stability. Importantly, the three characteristics of the sensors produced using the proposed platform all meet POC standards set by the Clinical and Laboratory Standards Institute (CLSI). This suggests possible approval of the biosensors for POC development. Furthermore, the detection range of the platform was demonstrated by constructing biosensors capable of detecting common POC targets, including circulating tumor cells (CTCs), DNA/RNA, and curcumin, and the devices were optimized for POC use. Overall, the platform developed in this study shows high potential for production of POC biosensors.


Assuntos
Técnicas Biossensoriais/normas , Técnicas Eletroquímicas/normas , Testes Imediatos/normas , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Curcumina/análise , DNA/análise , Eletrodos , Humanos , RNA/análise , Reprodutibilidade dos Testes
3.
Mikrochim Acta ; 187(1): 97, 2020 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-31907624

RESUMO

A stable and enzyme-free method is described for highly sensitive determination of telomerase activity. It is based on the use of a framework nucleic acid (FNA) nanomachine and doxorubicin-spherical nucleic acid (DSNA) tags. Upon incubation with telomerase, the primer-tetrahedron becomes elongated to form the handed swing arm. The extended swing arm autonomously moves along the predefined track consisting of entropy-tetrahedron by consecutive strand displacement under the aid of fuel-tetrahedron. As a result, many (entropy-tetrahedron)-(fuel-tetrahedron) complexes are assembled for combining the DSNA tags. This results in an amplified electrochemical signal, typically measured at around -0.63 V (Ag/AgCl). The use of an enzyme-free FNA nanomachine and of DSNA tags warrants outstandingly high stability and sensitivity. The method shows a broad dynamic correlation of telomerase activity in cell extracts. The analytical range extends from 10 to 1.0 × 104 HeLa cells mL-1 with a lower detection limit of 2 cells mL-1. The differences in telomerase activity between different cancer cells can be easily evaluated. The method was further verified by quantifying telomerase activity of cancer cells in accumulated normal cells. Therefore, the sensing method has great potential for clinical application. Graphical abstractSchematic representation of the electrochemical biosensor based on target induced framework nucleic acid nanomachine with doxorubicin-spherical nucleic acids (DSNA) tags, which can be used to the determination of telomerase activity in accumulated normal cells. dNTP: Deoxynucleotide triphosphates; FT: Fuel-tetrahedron.


Assuntos
Doxorrubicina/química , Técnicas Eletroquímicas/métodos , Ácidos Nucleicos/química , Telomerase/análise , Linhagem Celular Tumoral , Técnicas Eletroquímicas/normas , Células HeLa , Humanos , Nanomedicina/métodos , Neoplasias/diagnóstico , Neoplasias/enzimologia , Neoplasias/patologia , Telomerase/metabolismo
4.
Mikrochim Acta ; 187(1): 95, 2020 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-31903507

RESUMO

To increase the sensitivity of electrochemical sensor, Fe-MIL-88B-NH2 (Fe-MOF) with peroxidase-like activity is designed for the construction of immunoprobe. The Fe-MOF was prepared by one-step hydrothermalf method using 2-aminoterephthalic acid and iron(III) chloride. For the immunoprobe, it was fabricated by gold nanocomposite/Fe-MOF (Au/Fe-MOF) for the immobilization of labeling antibody (the antibody was used to conjuncting with label materials). The thin layer of Methylene Blue (MB) covered by reduced graphene oxide-gold nanocomposites (Au-rGO) serves as a substrate to covalently fix coating antibodies. The MB as a redox-active species was modified on the glass carbon electrode that can give a strong amperometric signal at 0.18 V (vs. Ag/AgCl). With the participation of H2O2, Fe-MOF can induce the Fenton reaction which degrades MB covered by Au-rGO on the substrate. The rest of MB on the surface of electrode becomes oxidized thereby generating a current signal. Square wave voltammetry (SWV) was used to quantify PSA. Under optimal conditions, the immunoassay is stable, specific and reproducible. It has a lower detection limit of 0.13 pg mL-1 (S/N = 3) and a wide analytical range that extends from 0.001 to 100 ng mL-1. Graphical abstractA sandwich-type amperometric immunoassay based on Fe-MOF-induced Fenton reaction was designed for sensitive determination of prostate specific antigen.


Assuntos
Técnicas Eletroquímicas/métodos , Calicreínas/análise , Estruturas Metalorgânicas/química , Nanocompostos/química , Peroxidase/metabolismo , Antígeno Prostático Específico/análise , Anticorpos Imobilizados/imunologia , Técnicas Eletroquímicas/normas , Eletrodos , Ouro , Humanos , Peróxido de Hidrogênio/química , Ferro , Calicreínas/imunologia , Azul de Metileno/química , Mimetismo Molecular , Oxirredução , Antígeno Prostático Específico/imunologia
5.
J Immunol Methods ; 470: 55-58, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31034878

RESUMO

FMS related tyrosine kinase 3 (FLT-3) is a tyrosine kinase expressed in early hematopoietic precursor cells and has roles in survival, proliferation, and differentiation. Bone marrow expression and mutagenic analysis of FLT-3 in Acute Myeloid Leukemia (AML) patients is well-characterized. However, the levels of circulating FLT-3 in serum have not been previously described. In this study we describe a quantitative electrochemiluminescent immunoassay that detects FLT-3 in human serum. Using this method we find that AML patients have elevated levels of circulating FLT-3 and these levels correlated to the percent blast counts in the bone marrow (BM).


Assuntos
Biomarcadores Tumorais/sangue , Técnicas Eletroquímicas/métodos , Imunoensaio/métodos , Leucemia Mieloide Aguda/diagnóstico , Medições Luminescentes/métodos , Tirosina Quinase 3 Semelhante a fms/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/genética , Medula Óssea/enzimologia , Medula Óssea/patologia , Técnicas Eletroquímicas/normas , Feminino , Expressão Gênica , Humanos , Imunoensaio/normas , Leucemia Mieloide Aguda/sangue , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/patologia , Luminescência , Medições Luminescentes/normas , Masculino , Pessoa de Meia-Idade , Variações Dependentes do Observador , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Tirosina Quinase 3 Semelhante a fms/genética
6.
Mikrochim Acta ; 186(2): 75, 2019 01 09.
Artigo em Inglês | MEDLINE | ID: mdl-30627835

RESUMO

An aptamer-based assay is described for the determination of trace levels of the cancer marker human epidermal growth factor receptor-2 (HER2) and living MCF-7 cells. The method is based on the use of a bimetallic MnFe Prussian blue analogue coupled to gold nanoparticles (MnFePBA@AuNP). Compared to pristine MnFe PBA nanocubes, the series of MnFePBA@AuNP exhibits a core-shell spherical nanostructure, and the shell thickness decreases from 99.9 nm down to 49.3 nm on increasing the fraction of AuNPs. The composite was placed on a gold electrode and incubated with the aptamer solution through electrostatic interaction. Then the modified electrode was employed to detect HER2 and MCF-7 cells using [Fe(CN)6]3-/4- as redox probe and displays good responses to both of them. Electrochemical impedance spectroscopy data show that the signal variation between each step during the whole procedure for the HER2 and MCF-7 cells detection can be embodied as the resistance value change between the [Fe(CN)6]3-/4- and electrode surface. The assay has a very low detection limit (0.247 pg∙mL-1) and works in the 0.001-1.0 ng∙mL-1 HER2 concentration range. It was also used to sense HER2 in MCF-7 cells, and this results in an assay that works within the 500-5 × 104 cell∙mL-1 cell concentration range and a 36 cell∙mL-1 detection limit. Furthermore, the aptamer-based assay is selective, acceptably reproducible, stable, and well feasible for the detection of HER2 and living MCF-7 cells in human serum. Graphical abstract Schematic of an electrochemical aptasensor based on the bimetallic MnFe Prussian blue analogue (MnFe PBA) coupling with gold nanoparticles (represented by MnFePBA@AuNPs). It was employed as the aptasensor for human epidermal growth factor receptor-2 (HER2), and living MCF-7 cells.


Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos , Ferrocianetos/química , Células MCF-7/citologia , Nanopartículas Metálicas/química , Receptor ErbB-2/análise , Contagem de Células , Impedância Elétrica , Técnicas Eletroquímicas/normas , Eletrodos , Ouro , Humanos , Ferro , Manganês
7.
J Mater Chem B ; 7(14): 2277-2283, 2019 04 14.
Artigo em Inglês | MEDLINE | ID: mdl-32254676

RESUMO

Herein, a light-driven, membrane-less and mediator-less self-powered cytosensing platform via integration of biofuel cells (BFCs) and a photoelectrochemical strategy was developed for ultrasensitive detection of circulating tumor cells (CTCs). To construct cytosensors, an elaborately designed SH-Sgc8c aptamer/AuNP/g-C3N4 photoelectrode was used as an alternative anode for glucose oxidation, avoiding the introduction of anodic enzymes. Initially, glucose could favorably reach the photoanode surface and be easily oxidized by the photogenerated holes, while the photogenerated electrons would transfer to the biocathode and achieve biocatalytic reduction of O2, leading to a high EOCV. However, in the presence of CTCs, they could preferentially interact with the Sgc8c aptamer via specific recognition, and then complexes with large steric hindrance were immobilized on the photoanode surface, which could greatly affect the electron transfer between glucose and the photoanode surface. In this case, the EOCV decreased sharply. Encouragingly, this self-powered cytosensor exhibited an ultrasensitive response to the target CTCs in a wide concentration range from 20 to 2 × 105 cells mL-1 with a low detection limit of 10 cells mL-1 (S/N = 3), being superior to those of the reported methods. Moreover, this as-proposed self-powered cytosensor integrated with a photoelectrochemical strategy possessed unique advantages of not requiring an external power source, being anodic enzyme-free, having a simple construction process, facile miniaturization, and high selectivity and sensitivity, providing a promising and powerful tool for fundamental biochemical research and clinical diagnosis.


Assuntos
Técnicas Biossensoriais/métodos , Contagem de Células/métodos , Técnicas Eletroquímicas/normas , Células Neoplásicas Circulantes/ultraestrutura , Fontes de Energia Bioelétrica , Técnicas Eletroquímicas/métodos , Células HeLa , Humanos
8.
Biosens Bioelectron ; 126: 230-239, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30439624

RESUMO

A novel label-free impedimetric immunosensor was fabricated for rapid, selective and sensitive detection and quantification of Cadherin-like protein 22 (CDH22), a cancer marker, in human serum by using easy and quickly prepared disposable ITO immunoelectrode. cancer marker, in human serum by using easy and quickly prepared disposable ITO immunoelectrode. The biosensing approach implied the use of ITO electrode coated with poly(phosphazene) polymer including benzaldehyde groups attached with CDH22 antibody and CDH22 antigens. Benzaldehyde side groups containing poly(phosphazene) film coated disposable ITO electrode were utilized as an immunosensing platform and anti-CDH22 antibodies bound to aldehyde groups of benzaldehyde substituted poly(phosphazene) (P-PHP) covalently. The immunosensor modification steps and affinity interaction between anti-CDH22 antibodies and CDH22 antigens were observed by EIS and CV in the presence of the redox couple. Furthermore, antibody immobilization was followed via FTIR and Raman spectroscopy. The morphological analyses of the suggested immunosensor during the fabrication steps were carried out with SEM and AFM monitoring. All the experimental parameters affecting the construction of the immunoelectrodes were optimized. The fabricated immunosensor exhibited an excellent working performance with a wide detection linear range (0.015-2.9 pg/mL) and low limit of detection (4.4 fg/mL). Moreover, the proposed immunosensor had great reproducibility, repeatability and long-term stability. Additionally, the fabricated immunosensor was successfully used in the quantification of CDH22 in human serum without any pretreatment.


Assuntos
Biomarcadores Tumorais/sangue , Técnicas Biossensoriais , Caderinas/sangue , Neoplasias/sangue , Anticorpos Imobilizados/química , Anticorpos Imobilizados/imunologia , Biomarcadores Tumorais/isolamento & purificação , Caderinas/isolamento & purificação , Técnicas Eletroquímicas/normas , Humanos , Imunoensaio/métodos , Limite de Detecção , Compostos Organofosforados/química , Polímeros/química
9.
Anal Bioanal Chem ; 411(1): 139-146, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30374725

RESUMO

A novel electrochemical luminescence (ECL) aptamer biosensor via polymerase amplification is constructed for label-free detection of leukemia marker mRNA (miR-16). In order to achieve the ultrasensitive detection of the target mRNA, the cyclic target chain displacement polymerization of leukemia marker mRNA assisted with Klenow fragment of DNA polymerase is employed. The determination is carried out by recording the ECL emission of pyridine ruthenium (Ru(bpy)32+) complexes embedded into the assistance DNA (ADNA) loaded on the nanogold surface, after the hybridization reaction between the probe DNA (PDNA) and the remaining sequence of the CP's stem part, and the formation of a core-shell sun-like structure. The mercapto-modified capture DNA (CP) is immobilized on the surface of a magneto-controlled glassy carbon electrode by Au-S bond. The CP is opened and hybridized with the target mRNA to form double-stranded DNA. In the presence of polymerase, primer DNA, and bases (dNTPs), the primer chain gets access to its complementary sequence of the stem part and then triggers a polymerization of the DNA strand, leading to the release of mRNA and starting the next polymerization cycle. Finally, the composite of PDNA-covered and ADNA-covered (embedded with Ru(bpy)32+) gold nanoparticles (hereafter called AuNPs@(PDNA+ADNA-Ru(bpy)32+) is added, and the ECL intensity is recorded. Because of the polymerization cycle and the aggregation of the illuminator of Ru(bpy)32+, the detected signal is amplified significantly. The results showed that the corresponding ECL signal has a good linear relationship with a logarithm of target mRNA concentration in the range of 1 × 10-16 to 1 × 10-7 mol/L, with a detection limit of 4.3 × 10-17 mol/L. The mRNA spiked in the human serum sample is determined, and the recoveries are from 97.2 to 102.0%. This sensor demonstrates good selectivity, stability, and reproducibility. Graphical abstract ᅟ.


Assuntos
Aptâmeros de Nucleotídeos/metabolismo , Biomarcadores Tumorais/sangue , Técnicas Biossensoriais/métodos , DNA Polimerase Dirigida por DNA/metabolismo , Técnicas Eletroquímicas/métodos , Leucemia/sangue , MicroRNAs/sangue , RNA Mensageiro/sangue , Calibragem , Sondas de DNA , Técnicas Eletroquímicas/normas , Eletrodos , Óxido Ferroso-Férrico/química , Ouro/química , Humanos , Limite de Detecção , Luminescência , Nanopartículas Metálicas/química , Reprodutibilidade dos Testes
10.
Mikrochim Acta ; 185(11): 520, 2018 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-30367266

RESUMO

A voltammetric sensor is described for the quantitation of propyl gallate (PG). A screen-printed carbon electrode (SPCE) was modified with reduced graphene sheets that were decorated with cobalt diselenide nanoparticles (CoSe2@rGO). The material was hydrothermally prepared and characterized by several spectroscopic techniques. The modified SPCE displays excellent electrocatalytic ability towards PG. Differential pulse voltammetry, with a peak voltage at 0.34 V (vs. Ag/AgCl) has a sensitivity of 12.84 µA·µM-1·cm-2 and a detection limit as low as 16 nM. The method is reproducible, selective, and practical. This method was applied to the determination of PG in spiked meat samples, and the result showed an adequate recovery. Graphical abstract Schematic of a new method for fast and sensitive electrochemical determination of the food additive propyl gallate in meat.


Assuntos
Cobalto/química , Técnicas Eletroquímicas/métodos , Carne/análise , Galato de Propila/análise , Selênio/química , Antioxidantes/análise , Técnicas Eletroquímicas/normas , Eletrodos , Aditivos Alimentares/análise , Grafite/química , Limite de Detecção , Óxidos/química
11.
J Pediatr Endocrinol Metab ; 31(6): 637-640, 2018 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-29750652

RESUMO

BACKGROUND: Bone metabolism involves many complex pathways that are disturbed by several bone diseases. The literature shows some limitations concerning pediatric reference intervals to bone markers, mainly because of the low number of patients included in the studies, the heterogeneity of methods, beyond the fact that it is time-consuming and expensive. The aim of this study was to determine reference values for ß-isomerized carboxy-terminal telopeptides collagen type I (ß-CTX), a marker of bone resorption, for children and adolescents. METHODS: Blood samples from 246 patients were collected and ß-CTX was measured using an electrochemiluminescence immunoassay (ECLI). RESULTS AND CONCLUSIONS: We propose reference ranges for ß-CTX concentration from the 2.5 percentile and 97.5 percentile for each age group. The reference values obtained, concerning children and adolescents, might be useful in the evaluation of diseases such as osteosarcoma and anorexia in both childhood as adolescence.


Assuntos
Biomarcadores/sangue , Reabsorção Óssea/diagnóstico , Colágeno Tipo I/sangue , Técnicas de Diagnóstico Endócrino/normas , Peptídeos/sangue , Adolescente , Fatores Etários , Reabsorção Óssea/sangue , Criança , Colágeno Tipo I/química , Técnicas Eletroquímicas/normas , Feminino , Humanos , Isomerismo , Medições Luminescentes/normas , Masculino , Peptídeos/química , Valores de Referência
12.
Respir Res ; 18(1): 103, 2017 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-28545517

RESUMO

BACKGROUND: Pleural effusions present a diagnostic challenge. Approximately 20% are associated with cancer and some 50% require invasive procedures to perform diagnosis. Determination of tumour markers may help to identify patients with malignant effusions. Two strategies are used to obtain high specificity in the differential diagnosis of malignant pleural effusions: a) high cut-off, and b) fluid/serum (F/S) ratio and low cut-off. The aim of this study is to compare these two strategies and to establish whether the identification of possible false positives using benign biomarkers - ADA, CRP and % of polymorphonuclear cells - improves diagnostic accuracy. METHODS: We studied 402 pleural effusions, 122 of them malignant. Benign biomarkers were determined in pleural fluid, and CEA, CA72-4, CA19-9 and CA15-3 in pleural fluid and serum. RESULTS: Establishing a cut-off value for each TM for a specificity of 100%, a joint sensitivity of 66.5% was obtained. With the F/S strategy and low cut-off points, sensitivity was 77% and specificity 98.2%, Subclassifying cases with negative benign biomarkers, both strategies achieved a specificity of 100%; sensitivity was 69.9% for single determination and 80.6% for F/S ratio. CONCLUSIONS: The best interpretation of TM in the differential diagnosis of malignant pleural effusions is obtained using the F/S ratio in the group with negative benign biomarkers.


Assuntos
Biomarcadores Tumorais/metabolismo , Derrame Pleural Maligno/diagnóstico , Derrame Pleural Maligno/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Diagnóstico Diferencial , Técnicas Eletroquímicas/normas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Derrame Pleural/diagnóstico , Derrame Pleural/metabolismo , Adulto Jovem
13.
Mikrochim Acta ; 185(1): 50, 2017 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-29594398

RESUMO

This article describes an impedimetric aptasensor for the prostate specific antigen (PSA), a widely accepted prostate cancer biomarker. A glassy carbon electrode (GCE) was modified with titanium oxide nanoparticles (TiO2) and silk fibroin nanofiber (SF) composite. The aptasensor was obtained by immobilizing a PSA-binding aptamer on the AuNP-modified with 6-mercapto-1-hexanol. The single fabrication steps were characterized by cyclic voltammetry and electrochemical impedance spectroscopy. The assay has two linear response ranges (from 2.5 fg.mL-1 to 25 pg.mL-1, and from 25 pg.mL-1 to 25 ng.mL-1) and a 0.8 fg.mL -1 detection limit. After optimization of experimental conditions, the sensor is highly selective for PSA over bovine serum albumin and lysozyme. It was successfully applied to the detection of PSA in spiked serum samples. Graphical abstract Schematic of the fabrication of an aptasensor for the prostate specific antigen (PSA). It is based on the use of a glassy carbon electrode modified with gold nanoparticles and titanium oxide-silk fibroin. The immobilization process of aptamer and interaction with PSA were followed by electrochemical impedance spectroscopy technique.


Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos , Antígeno Prostático Específico/análise , Neoplasias da Próstata/diagnóstico , Técnicas Eletroquímicas/normas , Eletrodos , Fibroínas , Humanos , Masculino , Nanopartículas Metálicas/química , Sondas Moleculares/química , Sondas Moleculares/normas , Nanofibras/química , Análise Espectral , Titânio
14.
Nanoscale ; 7(19): 9040-5, 2015 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-25920751

RESUMO

Graphene doped with heteroatoms can show new or improved properties as compared to the original undoped material. It has been reported that the type of heteroatoms and the doping conditions can have a strong influence on the electronic and electrochemical properties of the resulting material. Here, we wish to compare the electrochemical behavior of two n-type and two p-type doped graphenes, namely boron-doped graphenes and nitrogen-doped graphenes containing different amounts of heteroatoms. We show that the boron-doped graphene containing a higher amount of dopants provides the best electroanalytical performance in terms of calibration sensitivity, selectivity and linearity of response for the detection of gallic acid normally used as the standard probe for the quantification of antioxidant activity of food and beverages. Our findings demonstrate that the type and amount of heteroatoms used for the doping have a profound influence on the electrochemical detection of gallic acid rather than the structural properties of the materials such as amounts of defects, oxygen functionalities and surface area. This finding has a profound influence on the application of doped graphenes in the field of analytical chemistry.


Assuntos
Antioxidantes/análise , Técnicas Eletroquímicas , Grafite/química , Antioxidantes/normas , Boro/química , Calibragem , Técnicas Eletroquímicas/normas , Ácido Gálico/análise , Ácido Gálico/normas , Nitrogênio/química , Óxidos/química
15.
Analyst ; 139(22): 5740-6, 2014 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-25262699

RESUMO

We have described a highly sensitive method for detecting DNA hybridisation using a redox-labeled stem loop probe. The redox labels were poly(styrene-co-acrylic) (PSA) spheres of 454 nm diameter, modified by methylene blue (MB) deposited alternatively with poly(sodium 4-styrene sulphonate) (PSS) in a layer-by-layer process. Each PSA sphere carried approx. 3.7 × 10(5) molecules of MB, as determined optically. DIG-tagged stem loop probes were immobilised on screen printed electrodes bearing anti-DIG antibodies. Binding with the target enabled straightening of the stem loop, which made attachment to the MB-coated PSA spheres possible. For measuring the current from the direct reduction of MB by differential pulse voltammetry, a 30 mer DNA target common to 70 strains of Escherichia coli was calibrated across the range 1.0 fM to 100 pM (gradient = 3.2 × 10(-8) A (log fM)(-1), r(2) = 0.95, n = 60), with an LOD of ∼58 fM. By using Fe(CN)6(3-/4-) as a solution phase mediator for the MB reduction, we were able to lower the LOD to ∼39 aM (gradient = 5.95 × 10(-8) A (log aM)(-1), r(2) = 0.96, n = 30), which corresponds to the detection of 0.76 ag (∼50 molecules) in the 2 µL analyte sample. We hypothesise that the lowering of the LOD was due to the fact that not all the MB labels were able to contact the electrode surface.


Assuntos
DNA Bacteriano/química , Técnicas Eletroquímicas/normas , Eletrodos , Hibridização de Ácido Nucleico , Sequência de Bases , Calibragem , Escherichia coli/genética , Limite de Detecção , Oxirredução , Espectrofotometria Ultravioleta
16.
PLoS One ; 9(3): e92422, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24642754

RESUMO

Adenosine (ADO), a non-classical neurotransmitter and neuromodulator, and its metabolites adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP), have been shown to play an important role in a number of biochemical processes. Although their signaling is well described, it has been difficult to directly, accurately and simultaneously quantitate these purines in tissue or fluids. Here, we describe a novel method for measuring adenosine (ADO) and its metabolites using high performance liquid chromatography with electrochemical detection (HPLC-ECD). Using this chromatographic technique, we examined baseline levels of ADO and ATP, ADP and AMP in 6 different brain regions of the C57BL/6J mouse: stratum, cortex, hippocampus, olfactory bulb, substantia nigra and cerebellum and compared ADO levels in 5 different strains of mice (C57BL/6J, Swiss-Webster, FVB/NJ, 129P/J, and BALB/c). These studies demonstrate that baseline levels of purines vary significantly among the brain regions as well as between different mouse strains. These dissimilarities in purine concentrations may explain the variable phenotypes among background strains described in neurological disease models.


Assuntos
Adenosina/metabolismo , Encéfalo/metabolismo , Neurotransmissores/metabolismo , Adenosina/isolamento & purificação , Difosfato de Adenosina/isolamento & purificação , Difosfato de Adenosina/metabolismo , Monofosfato de Adenosina/isolamento & purificação , Monofosfato de Adenosina/metabolismo , Trifosfato de Adenosina/isolamento & purificação , Trifosfato de Adenosina/metabolismo , Animais , Cromatografia Líquida de Alta Pressão/normas , Técnicas Eletroquímicas/normas , Masculino , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Neurotransmissores/isolamento & purificação , Especificidade de Órgãos , Padrões de Referência
17.
Mater Sci Eng C Mater Biol Appl ; 33(8): 4527-32, 2013 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-24094155

RESUMO

In this paper, a graphene (GR) ionic liquid (IL) 1-octyl-3-methylimidazolium hexafluorophosphate and chitosan composite-modified carbon molecular wire electrode (CMWE) was fabricated by a drop-casting method and further applied to the sensitive electrochemical detection of adenosine-5'-monophosphate (AMP). CMWE was prepared with diphenylacetylene (DPA) as the modifier and the binder. The properties of modified electrode were examined by scanning electron microscopy, cyclic voltammetry and electrochemical impedance spectroscopy. Electrochemical behaviors of AMP was carefully investigated with enhanced responses appeared, which was due to the presence of GR-IL composite on the electrode surface with excellent electrocatalytic ability. A well-defined oxidation peak of AMP appeared at 1.314 V and the electrochemical parameters were calculated by electrochemical methods. Under the selected conditions, the oxidation peak current of AMP was proportional to its concentration in the range from 0.01 µM to 80.0 µM with the detection limit as 3.42 nM (3σ) by differential pulse voltammetry. The proposed method exhibited good selectivity and was applied to the detection of vidarabine monophosphate injection samples with satisfactory results.


Assuntos
Monofosfato de Adenosina/análise , Carbono/química , Quitosana/química , Técnicas Eletroquímicas , Grafite/química , Líquidos Iônicos/química , Monofosfato de Adenosina/normas , Calibragem , Catálise , Espectroscopia Dielétrica , Técnicas Eletroquímicas/normas , Eletrodos , Concentração de Íons de Hidrogênio , Imidazóis/química , Oxirredução , Propriedades de Superfície
18.
Anal Chem ; 85(16): 7654-8, 2013 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-23919317

RESUMO

Direct electrochemical measurements of biological events are often challenging because of the low signal relative to the magnitude of the background and noise. When choosing a data processing approach, the frequency and phase content of the data must be considered. Here, we employ a zero-phase (infinite impulse response (IIR)) filter to remove the noise from the analytical signal, while preserving the phase content. In fast-scan cyclic voltammetry, the frequency content of the signal is a function of the scan rate of the applied waveform. Fourier analysis was used to develop a relationship between scan rate and the filter cutoff frequency to maximize the reduction in noise, while not altering the true nature of the analytical signal. The zero-phase filter has the same effect as traditional filters with regards to increasing the signal-to-noise ratio. Because the zero-phase filter does not introduce a change to ΔEpeak, the heterogeneous electron rate transfer constant (0.10 cm/s) for ferrocene is calculated accurately. The zero-phase filter also improves electrochemical analysis of signaling molecules that have their oxidation potential close to the switching potential. Lastly, a quantitative approach to filtering amperometric traces of exocytosis based on the rise time was developed.


Assuntos
Coleta de Dados , Técnicas Eletroquímicas/normas , Processamento de Sinais Assistido por Computador , Animais , Exocitose , Análise de Fourier , Cinética , Células PC12 , Ratos
19.
Anal Chim Acta ; 786: 34-8, 2013 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-23790289

RESUMO

In this communication, the first nonelectrocatalysis-type hydrogen peroxide electrochemical sensor is reported. The electroactive iron(III) diethylenetriaminepentaacetic acid (DTPA-Fe(III)) complex is immobilized on the cysteamine (cys) modified nanoporous gold (NPG) films by covalent method. The immobilized DTPA-Fe(III) complex quickly communicates an electron with the electrode. Upon addition of hydrogen peroxide, however, hydrogen peroxide inhibits the direct electron transfer of the DTPA-Fe(III) complex due to the generation of nonelectroactive DTPA-Fe(III)-H2O2 complex. Based on quenching mechanism, the first hydrogen peroxide electrochemical sensor based on a nonelectrocatalytic mechanism is developed. The novel hydrogen peroxide electrochemical sensor has the ultralow detection limit (1.0×10(-14) M) and wide linear range (1.0×10(-13) to 1.0×10(-8) M) with excellent reproducibility and stability.


Assuntos
Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Compostos Férricos/química , Peróxido de Hidrogênio/química , Técnicas Biossensoriais/normas , Técnicas Eletroquímicas/normas
20.
Anal Chim Acta ; 772: 26-32, 2013 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-23540244

RESUMO

The electrochemical detection of cell lines of MCF-7 (human breast cancer) has been reported, using magnetic beads for the separation tool and high-affinity DNA aptamers for signal recognition. The high specificity was obtained by using the magnetic beads and aptamers, and the good sensitivity was realized with the signal amplification of DNA capped CdS or PbS nanocrystals. The ASV (anodic stripping voltammetry) technology was employed for the detection of cadmic cation and lead ions, for electrochemical assay of the amount of the target cells and biomarkers on the membrane of target cells, respectively. This electrochemical method could respond to as low as 100 cells mL(-1) of cancer cells with a linear calibration range from 1.0×10(2) to 1.0×10(6) cells mL(-1), showing very high sensitivity. Moreover, the amounts of HER-3 which were overexpressed on MCF-7 cells were calculated correspond to be 3.56×10(4) anti-HER-3 antibody molecules. In addition, the assay was able to differentiate between different types of target and control cells based on the aptamers and magnetic beads used in the assay, indicating the wide applicability of the assay for early and accurate diagnose of cancers.


Assuntos
Antígenos de Neoplasias/análise , Aptâmeros de Nucleotídeos/química , Separação Celular/métodos , Técnicas Eletroquímicas/métodos , Nanopartículas Metálicas/química , Receptor ErbB-3/análise , Antígenos de Neoplasias/química , Compostos de Cádmio/química , Linhagem Celular Tumoral , Separação Celular/normas , Técnicas Eletroquímicas/normas , Humanos , Chumbo/química , Imãs , Receptor ErbB-3/química , Sensibilidade e Especificidade , Sulfetos/química
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