RESUMO
Hydroquinone (HQ) is one of the major deleterious metabolites of benzene in the human body, which has been implicated to cause various human diseases. In order to fabricate a feasible sensor for the accurate detection of HQ, we attempted to electrochemically modify a piece of common 2B pencil lead (PL) with the conductive poly(3,4-ethylenedioxythiophene) or PEDOT film to construct a PEDOT/PL electrode. We then examined the performance of PEDOT/PL in the detection of hydroquinone with different voltammetry methods. Our results have demonstrated that PEDOT film was able to dramatically enhance the electrochemical response of pencil lead electrode to hydroquinone and exhibited a good linear correlation between anodic peak current and the concentration of hydroquinone by either cyclic voltammetry or linear sweep voltammetry. The influences of PEDOT film thickness, sample pH, voltammetry scan rate, and possible chemical interferences on the measurement of hydroquinone have been discussed. The PEDOT film was further characterized by SEM with EDS and FTIR spectrum, as well as for stability with multiple measurements. Our results have demonstrated that the PEDOT modified PL electrode could be an attractive option to easily fabricate an economical sensor and provide an accurate and stable approach to monitoring various chemicals and biomolecules.
RESUMO
Dopamine (DA) is an important neurotransmitter responsible for the functions and activities of multiple systems in human. Electrochemical detection of DA has the advantages of fast analysis and cost-effectiveness, while a regular electrode probe is restricted to laboratory use because the probe size is too large to be suitable for an in vivo or in vitro analysis. In this study, we have developed porphyrin-based metal organic framework (MOF525) and poly(3,4-ethylenedioxythiophene) (PEDOT)-based composites to modify microelectrode for DA detection. Two types of PEDOT monomers with different functional groups were investigated in this study. By varying the monomer ratios, electrolyte concentrations, and electropolymerization temperature, it was found that the PEDOT monomer containing carboxylic group facilitated the formation of regular morphology during the electropolymerization process. The uniform morphology of the PEDOT promoted the electron transmission efficiency in the same direction, while the MOF525 provided a large reactive surface area for electrocatalysis of DA. Thus, the MOF525/PEDOT composite improved the sensitivity-to-noise ratio of DA signaling, where the sensitivity reached 11 nA/µM in a good linear range of 4-100 µM. In addition, porphyrin-based MOF could also increase the selectivity to DA against other common clinical interferences, such as ascorbic acid and uric acid. The as-synthesized microelectrode modified with MOF525/PEDOT in this study exhibited great potential in real time analysis.
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This study demonstrates a new, robust, and accessible deposition technique of metal nanoparticle arrays (NPAs), which uses nanoporous anodic alumina (NAA) as a template for capillary force-assisted convective colloid (40, 60, and 80 nm diameter Au) assembly. The NPA density and nanoparticle size can be independently tuned by the anodization conditions and colloid synthesis protocols. This enables production of non-touching variable-density NPAs with controllable gaps in the 20-60 nm range. The NPA nearest neighbor center distance in the present study was fixed to 100 nm by the choice of anodization protocol. The obtained Au NPAs have the resonant scattering maxima in the visible spectral range, with a refractometric sensitivity, which can be tuned by the variation of the array density. The thickness of the NAA layer in an Aluminum-NAA-NPA multilayer system enables further tuning of the resonance frequency and optimization for use with specific molecules, e.g., to avoid absorption bands. Applicability of the mentioned multilayers for colorimetric refractive index (RI) sensing is demonstrated. Their use as Surface-Enhanced Raman Scattering (SERS) substrates is tested using hemoglobin as a biological probe molecule.
RESUMO
The general clinical procedure for viral DNA detection or gene mutation diagnosis following polymerase chain reaction (PCR) often involves gel electrophoresis and DNA sequencing, which is usually time-consuming. In this study, we have proposed a facile strategy to construct a DNA biosensor, in which the platinum electrode was modified with a dual-film of electrochemically synthesized poly(3,4-ethylenedioxythiophene) (PEDOT) resulting in immobilized gold nanoparticles, with the gold nanoparticles easily immobilized in a uniform distribution. The DNA probe labeled with a SH group was then assembled to the fabricated electrode and employed to capture the target DNA based on the complementary sequence. The hybridization efficiency was evaluated with differential pulse voltammetry (DPV) in the presence of daunorubicin hydrochloride. Our results demonstrated that the peak current in DPV exhibited a linear correlation the concentration of target DNA that was complementary to the probe DNA. Moreover, the electrode could be reused by heating denaturation and re-hybridization, which only brought slight signal decay. In addition, the addition of the oxidized form of nicotinamide adenine dinucleotide (NADâº) could dramatically enhance the sensitivity by more than 5.45-fold, and the limit-of-detection reached about 100 pM.
Assuntos
Técnicas Biossensoriais/métodos , Compostos Bicíclicos Heterocíclicos com Pontes/química , DNA/análise , Condutividade Elétrica , Polímeros/química , Temperatura , Técnicas Eletroquímicas , Eletrodos , Ouro , Nanopartículas Metálicas , Platina/química , Poliestirenos/químicaRESUMO
In this study, we provided the detailed characterizations of our recent HRP-AuNPs/PEDOT:BSA/Pt biosensor, constructed through a simple fabrication procedure with improved stability and good sensitivity. Raman and Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy not only confirmed the synthesis of conductive PEDOT where BSA was the template for the polymerization, but also provided further insights into the stable immobilization of AuNP on the PEDOT:BSA film. Scanning electron microscopy revealed that the attachment of AuNPs were stable under a high salt environment. The current technology demonstrates a feasible procedure to form a functional AuNPs/PEDOT:BSA film that has potential applications in the fabrication of various biosensors and electric devices.
Assuntos
Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Peroxidase do Rábano Silvestre/metabolismo , Animais , Técnicas Biossensoriais/instrumentação , Compostos Bicíclicos Heterocíclicos com Pontes/química , Bovinos , Eletrodos , Ouro/química , Peróxido de Hidrogênio/análise , Nanopartículas Metálicas/química , Microscopia Eletrônica de Transmissão , Espectroscopia Fotoeletrônica , Platina/química , Polímeros/química , Soroalbumina Bovina/química , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
In this study, we have investigated the contribution of bovine serum albumin (BSA) to the durability of the electrochemically synthesized poly(3,4-ethylenedioxythiophene) (PEDOT) film on a platinum (Pt) electrode. The electrode was capable to effectively adsorb the nano Au particles (AuNPs) to form a uniform layout, which was then able to immobilize the horseradish peroxidase (HRP) to construct a functional HRP/AuNPs/PEDOT(BSA)/Pt biosensor. Cyclic voltammetry was employed to evaluate the performance of the biosensor through the measurement of hydrogen peroxide. Our results revealed a satisfied linear correlation between the cathodic current and the concentration of H2O2. Furthermore, the addition of oxidized form of nicotinamide adenine dinucleotide, or NADâº, as the electron transfer mediator in the detection solution could dramatically enhance the sensitivity of detection by about 35.5%. The main advantages of the current biosensor are its durability, sensitivity, reliability, and biocompatibility.
Assuntos
Técnicas Biossensoriais/métodos , Peróxido de Hidrogênio/isolamento & purificação , Nanopartículas/química , Soroalbumina Bovina/química , Animais , Compostos Bicíclicos Heterocíclicos com Pontes/química , Bovinos , Eletrodos , Enzimas Imobilizadas/química , Ouro/química , Peroxidase do Rábano Silvestre/química , Polímeros/químicaRESUMO
In this work, hydroxyapatite (HA) mineralized on chitosan (CS)-coated poly(lactic acid) (PLA) nanofiber mat was prepared and compared in terms of mineralization characteristics. Significant calcium phosphate crystals formed on various concentrations of CS-coated PLA fiber mat with better uniformity after 2h of incubation in 10 times simulated body fluid (10× SBF). X-ray diffraction results further indicated that the composition of the deposited mineral was a mixture of dicalcium phosphate dehydrates and apatite. Chitosan, a cationic polysaccharide, can promote more nucleation and growth of calcium phosphate under conditions of 0.4% chitosan concentrations. These results indicated that HA-mineralized on CS-coated PLA fiber mat can be prepared directly via simply using CS coating followed by SBF immersion, and the results also suggest that this composite can mimic structural, compositional, and biological functions of native bone and can serve as a good candidate for bone tissue engineering (BTE).
Assuntos
Fosfatos de Cálcio/química , Quitosana/farmacologia , Materiais Revestidos Biocompatíveis/farmacologia , Ácido Láctico/farmacologia , Minerais/química , Nanofibras/química , Polímeros/farmacologia , Engenharia Tecidual/métodos , Fosfatase Alcalina/metabolismo , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Cristalização , Durapatita/farmacologia , Ensaios Enzimáticos , Humanos , Nanofibras/ultraestrutura , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/enzimologia , Osteoblastos/ultraestrutura , Poliésteres , Espectrometria por Raios X , Espectroscopia de Infravermelho com Transformada de Fourier , Análise Espectral Raman , Difração de Raios XRESUMO
The conductive polymer, poly(3,4-ethylenedioxythiophene) (PEDOT), possesses attractive properties that show the potential applications in many fields. In this study, we have proposed a two-stage enzymatic synthesis of conductive PEDOT. Horseradish peroxidase (HRP) acts as the catalyst to promote the generation of EDOT free radicals followed by the polymerization under the room temperature in the presence of poly(sodium 4-styrenesulfonate) (PSS), then a mild heating process is employed for further chain extension. The final PEDOT:PSS is purified with n-butanol and subjected to various characterizations, which indicate that PEDOT with enzymatic approach exhibits a similar molecular structure to that with chemical method. However, the enzymatically synthesized PEDOT:PSS demonstrates advantages, such as stable integration of PEDOT with PSS and better electrochemical properties, suggesting its future prospective applications.
Assuntos
Compostos Bicíclicos Heterocíclicos com Pontes/metabolismo , Polímeros/metabolismo , Biotecnologia , Compostos Bicíclicos Heterocíclicos com Pontes/química , Catálise , Condutividade Elétrica , Técnicas Eletroquímicas , Peroxidase do Rábano Silvestre/metabolismo , Temperatura Alta , Estrutura Molecular , Polímeros/química , Poliestirenos/química , Poliestirenos/metabolismo , Espectrofotometria , Espectroscopia de Infravermelho com Transformada de Fourier , Análise Espectral RamanRESUMO
In this study, we have fabricated a label free DNA biosensor by modifying the platinum wire with electrochemically synthesized poly(3,4-ethylenedioxythiopene) and poly(p-aminobenzoic acid). A designed single-strand DNA oligo was immobilized with the carboxyl group of poly(p-aminobenzoic acid) and served as the probe, a target DNA was then hybridized with the probe under a proper condition. Differential pulse voltammetry was performed to characterize the hybridization efficiency in the presence of daunorubicin hydrochloride that was able to be intercalated into the hybridized double-strand DNA and possessed the redox activity. Our results revealed a satisfied linear correlation between the peak current of differential pulse voltammetry and the concentration of complementary target DNA. On the other hand, the mismatches between the target- and probe-DNA caused a significant reduction of electrochemical response, in which was correlated with the amount of mismatched base pairs, therefore the current DNA biosensor had potential applications not only in DNA quantification but also in mutation detection for clinical diagnostics and laboratory applications.
Assuntos
Ácido 4-Aminobenzoico/química , Técnicas Biossensoriais/instrumentação , Compostos Bicíclicos Heterocíclicos com Pontes/química , Condutometria/instrumentação , Análise Mutacional de DNA/instrumentação , Sondas de DNA/genética , Platina/química , Polímeros/química , Eletrodos , Desenho de Equipamento , Análise de Falha de Equipamento , Estudos de Viabilidade , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Polyaniline (PANI), an attractive conductive polymer, has been successfully applied in fabricating various types of enzyme-based biosensors. In this study, we have employed mesoporous silica SBA-15 to stably entrap horseradish peroxidase (HRP), and then deposited the loaded SBA-15 on the PANI modified platinum electrode to construct a GA/SBA-15(HRP)/PANI/Pt biosensor. The mesoporous structures and morphologies of SBA-15 with or without HRP were characterized. Enzymatic protein assays were employed to evaluate HRP immobilization efficiency. Our results demonstrated that the constructed biosensor displayed a fine linear correlation between cathodic response and H(2)O(2) concentration in the range of 0.02 to 18.5 mM, with enhanced sensitivity. In particular, the current approach provided the PANI modified biosensor with improved stability for multiple measurements.
RESUMO
Polyaniline (PANI) is considered as one of the most fascinating conductive polymers in fabricating enzyme-based biosensors. Nevertheless, to improve both sensitivity and stability of the PANI-modified biosensor has been and continues to be a technical challenge. In this study, we have electrochemically synthesized the PANI film on a platinum (Pt) electrode and then used this electrode to construct a horseradish peroxidase (HRP)-based biosensor for the detection of hydrogen peroxide (H(2)O(2)). The electrochemical and structural properties of electrodes were characterized with scanning electron microscopy (SEM), thermogravimetric analysis (TGA), Fourier-transform infrared (FTIR) spectrophotometer, and cyclic voltammetry (CV). It was interestingly found that the PANI film synthesized in the presence of bovine serum albumin (BSA) has provided the electrode with enhanced sensitivity and excellent stability. Our results suggested that the embedded BSA might serve as an initial template for aniline polymerization and stabilized the microstructure of the PANI film significantly. The constructed HRP/PANI(BSA)/Pt electrode also exhibited a fine linear correlation with H(2)O(2) concentration. This approach by implanted BSA was useful for improving the sensitivity and stability of PANI-modified biosensor.
Assuntos
Técnicas Biossensoriais/instrumentação , Eletrodos , Soroalbumina Bovina/química , Compostos de Anilina/química , Técnicas Biossensoriais/métodos , Peroxidase do Rábano Silvestre/química , Platina/química , Sensibilidade e Especificidade , Espectroscopia de Infravermelho com Transformada de Fourier , TermogravimetriaRESUMO
Polyaniline (PANI) has been shown to possess excellent catalytic activity toward oxygen reduction, however, this molecule may interfere with the electrochemical measurement of other targets when using a polyaniline modified platinum (PANI/Pt) electrode. In this study, we have demonstrated the considerable effects of dissolved oxygen on the sensing of hydrogen peroxide with the PANI/Pt electrode. Accordingly, we proposed a strategy to eliminate the influence of dissolved oxygen with oxygen scavengers. Our results indicated that as an oxygen scavenger sodium thiosulfate was very effective in the removal of dissolved oxygen from the sample solution, and had negligible effect on the quantification of hydrogen peroxide when its applied concentration was below 1 mM.
RESUMO
Adenines mismatched with guanines or 7,8-dihydro-8-oxo-deoxyguanines that arise through DNA replication errors can be repaired by either base excision repair or mismatch repair. The human MutY homolog (hMYH), a DNA glycosylase, removes adenines from these mismatches. Human MutS homologs, hMSH2/hMSH6 (hMutSalpha), bind to the mismatches and initiate the repair on the daughter DNA strands. Human MYH is physically associated with hMSH2/hMSH6 via the hMSH6 subunit. The interaction of hMutSalpha and hMYH is not observed in several mismatch repair-defective cell lines. The hMutSalpha binding site is mapped to amino acid residues 232-254 of hMYH, a region conserved in the MutY family. Moreover, the binding and glycosylase activities of hMYH with an A/7,8-dihydro-8-oxo-deoxyguanine mismatch are enhanced by hMutSalpha. These results suggest that protein-protein interactions may be a means by which hMYH repair and mismatch repair cooperate in reducing replicative errors caused by oxidized bases.