RESUMO
Technetium (99Tc) is a highly toxic radioactive nuclear wastewater contaminant. Real-time detection of 99Tc is very difficult due to its difficult-to-complex nature. Herein, a novel three-dimensional ionic olefin-linked conjugated microporous polymer (TFPM-EP-Br) is constructed using tetrakis(4-aldehyde phenyl)methane (TFPM) as the central monomer. The unique cationic cavity and highly hydrophobic framework enable TFPM-EP-Br to act as a fluorescent sensor for TcO4-. The fluorophores of TFPM-EP-Br can be quenched due to electron transfer from TFPM-EP-Br to TcO4- and the formation of strongly nonfluorescent complexes. Meanwhile, the regular pore channels are beneficial for the fast mass transfer of TcO4-, resulting in an ultrafast response time (less than 2 s) with an ultralow detection limit (33.3 nM). In addition, the ultrahigh specific surface area enables TFPM-EP-Br to combine the ability to synergistically detect and remove radioactive 99Tc. From this perspective, the novel conjugated microporous polymer has made a breakthrough in the detection and extraction of radioactive contaminants.
Assuntos
Polímeros , Águas Residuárias , Alcenos , Cátions , Tecnécio/químicaRESUMO
The traditional detection of telomerase activity is mainly based on the polymerase chain reaction (PCR), which has the disadvantages of being time-consuming and susceptible to interferences; thus, here, we propose a facile method for the fabrication of fluorescent tungsten oxide quantum dots (WOx QDs) and employ them for telomerase activity sensing. It is found that the fluorescence of WOx QDs can be significantly quenched by hemin based on the inner filter effect (IFE). However, in the presence of telomerase, the primer-DNA can be extended to generate repeating units of TTAGGG to form G-quadruplex and thus, hemin can be encapsulated to reduce its absorbance, resulting in decreased IFE and efficient fluorescence recovery of WOx QDs. Based on the fluorescence changes of IFE between hemin and WOx QDs, the telomerase activity within the range of 50-30 000 HeLa cells can be detected and the lowest detection amount can reach 17 cells. The method exhibits good versatility that can also be applied to telomerase detection in A549 and L929 cells. In addition, because of the good biocompatibility of the sensor, it can be used for the real-time monitoring of telomerase activity in living cells, thus showing great potential in tumor diagnosis and inhibitor drug screening.
Assuntos
Técnicas Biossensoriais/métodos , Óxidos/química , Pontos Quânticos/química , Telomerase/metabolismo , Tungstênio/química , Animais , Materiais Biocompatíveis/síntese química , Materiais Biocompatíveis/química , Linhagem Celular , DNA/química , DNA/metabolismo , Primers do DNA/metabolismo , Quadruplex G , Hemina/química , Humanos , Camundongos , Microscopia Confocal , Espectrometria de FluorescênciaRESUMO
A fluorometric method is described for the determination of thrombin. Polymer nanoparticles containing the luminol-terbium(III) complex (luminol-Tb) were prepared where luminol acts as the bridging ligand, and Tb(III) acts as the central metal ion. Thrombin possesses a large number of electrons donating groups that coordinate with luminol-Tb. Following coordination, the rigidity of the linker is increased, and this decreases the non-radiative decay rate and induces an increase in fluorescence intensity at 430 nm. Hence, thrombin can be fluorometrically determined. The detection limit of thrombin is as low as 3.5 pM (at an SNR of 3). This is about 10 times lower than assays using an aptamer. The method was applied in the determination of thrombin in human serum via the standard addition method and gave satisfying results. Graphical abstractSchematic representation of the preparation of the luminol-Tb(III) complex in a nanoparticle host by the self-assembly of luminol and Tb(III) ions. Thrombin readily coordinates with the luminol-Tb(III) system, and this results in particle aggregation. The blue fluorescence of luminol increases strongly, and this effect provides the basis for fluorometric determination of thrombin.
Assuntos
Complexos de Coordenação/química , Fluorescência , Nanopartículas/química , Polímeros/química , Térbio/química , Trombina/análise , Complexos de Coordenação/síntese química , Luminol/química , Estrutura Molecular , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
In this work, a simple and rapid approach was developed for separation and detection of chiral compounds based on a magnetic molecularly imprinted polymer modified poly(dimethylsiloxane) (PDMS) microchip coupled with electrochemical detection. Molecularly imprinted polymers were prepared employing Fe3 O4 nanoparticles (NPs) as the supporting substrate and norepinephrine as the functional monomer in the presence of template molecule in a weak alkaline solution. After extracting the embedded template molecules, Fe3 O4 @polynorepinephrine NPs (MIP-Fe3 O4 @PNE NPs) showed specific molecular recognition selectivity and high affinity towards the template molecule, which were then used as stationary phase of microchip capillary electrochromatography for chiral compounds separation. Mandelic acid and histidine enantiomers were used as model compounds to test the chiral stationary phase. By using R-mandelic acid as the template molecule, mandelic acid enantiomer was effectively separated and detected on the MIP-Fe3 O4 @PNE NPs modified PDMS microchip. Moreover, the successful separation of histidine enantiomers on the MIP-Fe3 O4 @PNE NPs modified microchip using L-histidine as template molecule was also achieved.
Assuntos
Eletrocromatografia Capilar/métodos , Nanopartículas de Magnetita/química , Impressão Molecular/métodos , Eletro-Osmose , Histidina/química , Limite de Detecção , Ácidos Mandélicos/química , Norepinefrina/química , Polímeros/química , Reprodutibilidade dos Testes , EstereoisomerismoRESUMO
Lanthanide coordination polymer nanoparticles (Ln-CPNs) have been recently demonstrated as excellent platforms for biomolecule detection. In this work, we synthesized novel cerium coordination polymer nanoparticles ATP-Ce-Tris CPNs in a simple and quick way using ATP molecules as the biocompatible ligands to Ce(3+) ions in tris(hydroxymethyl)aminomethane hydrochloric (Tris-HCl) solution. In view of the excellent free radical scavenging property of cerium compounds, which is ascribed to the mixed valence state (Ce(3+), Ce(4+)) and the reversible switch from Ce(3+) to Ce(4+), the synthesized ATP-Ce-Tris CPNs was used as artificial peroxidase to selectively and sensitively detect H2O2. The sensing mechanism depends on the oxidation of the fluorescent ATP-Ce(III)-Tris CPNs to nonfluorescent ATP-Ce(IV)-Tris CPNs by H2O2. Compared with those inorganic cerium oxide sensors, this kind of fluoresence ATP-Ce-Tris CPNs sensor needs no additional organic redox dye, such as ABTS (2,20-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), TMB (3,3,5,5-tetramethylbenzidine), or fluorescein as signal molecules. Moreover, such ATP-Ce-Tris CPNs sensor exhibited a more sensitive response to H2O2 with a detection limit down to 0.6 nM, which is 2 orders of magnitude lower than those of cerium oxide sensors. This sensing platform was further extended to the detection of glucose in combination with the specific catalytic effect of glucose oxidase (GOx) for the oxidation of glucose and formation of H2O2.
Assuntos
Técnicas Biossensoriais/métodos , Peróxido de Hidrogênio/análise , Elementos da Série dos Lantanídeos/química , Nanopartículas Metálicas/química , Polímeros/química , Trifosfato de Adenosina/química , Catálise , Cério/química , Glucose/análise , Glucose Oxidase/metabolismo , Limite de Detecção , Peroxidase/química , Peroxidase/metabolismo , Espectrometria de FluorescênciaRESUMO
In this paper, using the self-polymerization of norepinephrine (NE) and its favorable film-forming property, a simple and green preparation approach was developed to modify a PDMS channel for enantioseparation of chiral compounds. After the PDMS microchip was filled with NE solution, poly(norepinephrine) (PNE) film was gradually formed and deposited on the inner wall of microchannel as permanent coating via the oxidation of NE by the oxygen dissolved in the solution. Due to possessing plentiful catechol and amine functional groups, the PNE-coated PDMS microchip exhibited much better wettability, more stable and suppressed EOF, and less nonspecific adsorption. The water contact angle and EOF of PNE-coated PDMS substrate were measured to be 13° and 1.68 × 10(-4) cm(2) V(-1) s(-1) , compared to those of 108° and 2.24 × 10(-4) cm(2) V(-1) s(-1) from the untreated one, respectively. Different kinds of chiral compounds, such as amino acid enantiomer, drug enantiomer, and peptide enantiomer were efficiently separated utilizing a separation length of 37 mm coupled with in-column amperometric detection on the PNE-coated PDMS microchips. This facile mussel-inspired PNE-based microchip system exhibited strong recognition ability, high-performance, admirable reproducibility, and stability, which may have potential use in the complex biological analysis.
Assuntos
Eletroforese em Microchip/métodos , Norepinefrina/metabolismo , Animais , Bivalves , Dimetilpolisiloxanos , Eletroforese em Microchip/instrumentação , Química Verde , Polimerização , Reprodutibilidade dos Testes , EstereoisomerismoRESUMO
To ensure the long-term sustainable development of nuclear energy as well as the prevention and control of uranium pollution, new materials that can simultaneously detect and separate uranium are still urgently needed. Herein, a new fluorescent covalent organic polymer (COP), namely HT-COP-AO, was synthesized andemployed as both the fluorescent probe and absorbent for simultaneous uranium detection and separationconsidering its excellent fluorescence property and strong uranium coordination ability. The results showed that the fluorescence of HT-COP-AO was quickly quenched by uranium within 2 min, and the limit of detection was 0.23 µM (3σ/K). Further studies implied that uranium was coordinated with the amidoxime groups of HT-COP-AO through U-N and O = U = O bonds, which resulted in electron transfer from uranium to HT-COP-AO and quenching the fluorescence of HT-COP-AO consequently. Meanwhile, HT-COP-AO exhibited excellent absorption ability towards uranium, and the maximum absorption capacity (qmax = 401.3 mg/g) was higher than most reported amidoxime modified materials. The HT-COP-AO also showed high selectivity for both uranium detection and separation which makes it a great promising for uranium monitoring in real water samples.
Assuntos
Urânio , Corantes Fluorescentes , Transporte de Elétrons , PolímerosRESUMO
The elimination of anion is of great importance from radioactive nuclear waste containing 99TcO4- by rationally designing anion-scavenging materials with high density of charge and more accessible adsorption sites. Herein, a tailor-made cationic organic polymer with donor-acceptor (D-A) structure, namely TrDCPN, was successfully synthesized by rationally modifying the benzimidazole unit for efficient trapping the perrhenate (ReO4-) as a 99Tc surrogate. Systematic control of the skeleton affect enables the material to integrate a variety of features, surmounting the long-term challenge of 99TcO4-/ReO4- remediation under extreme conditions of high acid/base and high ionic strength. Furthermore, the TrDCPN shows excellent affinity toward ReO4- in the existence of large excess of competitive anions (SO42-, NO3- and PO43-etc.) as well as promising reusability for trapping ReO4-. The excellent stability and separation were derived from the introduction of large conjugated modules, triazine core and hydrophobic. More importantly, the synthetic cationic organic polymer with D-A feature was first proved that the introduction of halogen can effectively enhance the backbone charge, and increase the adsorption capacity by synergy of ion exchange, electrostatic interaction and δ hole-anion interaction. The adsorption capacity of TrDCPN can be up to 420.3 mg/g and reach equilibrium within 20 min. It is noteworthy that TrDCPN successfully immobilizes ReO4- from simulated Hanford waste with a high separation efficiency of 93 %, providing a new paradigm for material design to dispose of the problem of radioactive pollutants in the environment.
Assuntos
Halogênios , Resíduos Radioativos , Polímeros , Cátions , Adsorção , Troca IônicaRESUMO
In this paper, a novel, simple, economical and environmentally friendly method based on in situ chemically induced synthesis strategy was designed and developed for the modification of a poly(dimethylsiloxane) (PDMS) microchip channel with polydopamine/gold nanoparticles (PDA/Au NPs) to create a hydrophilic and biofouling resistant surface. Dopamine as a reductant and a monomer, and HAuCl(4) as an oxidant to trigger dopamine polymerization and the source of metallic nanoparticles, were filled into the PDMS microchannel to yield in situ a well-distributed and robust PDA/Au NP coating. Au NPs were highly and uniformly dispersed in/on the PDA matrix with a narrow size distribution, as verified by scanning electron microscopy and UV-vis spectra. Compared with the native PDMS microchannel, the modified surfaces exhibited much better wettability, high stability and suppressed electroosmotic mobility, and less nonspecific adsorption towards biomolecules. The water contact angle and EOF of PDA/Au NP-coated PDMS microchip were measured to be 13° and 4.17×10(-4) cm(2)/V s, compared to those of 111° and 5.33×10(-4) cm(2)/V s from the native one, respectively. Fast and efficient separations of five amino acids such as arginine, proline, histidine, valine and threonine suggested greatly improved electrophoretic performance of the PDA/Au NP-functionalized PDMS microchips. This one-step procedure offers an effective approach for a biomimetic surface design on microfluidic chips, which is promising in high-throughput and complex biological analysis.
Assuntos
Aminoácidos/isolamento & purificação , Dimetilpolisiloxanos/química , Eletroforese em Microchip/instrumentação , Ouro/química , Nanopartículas Metálicas/química , Eletro-Osmose , Eletroforese em Microchip/métodos , Concentração de Íons de Hidrogênio , Indóis , Cinética , Microscopia Eletrônica de Varredura , Polímeros , Reprodutibilidade dos Testes , Difração de Raios XRESUMO
In this paper, titanium dioxide nanoparticles (TiO(2) NPs) were employed to construct a functional film on PDMS microfluidic channel surface, which was formed by sequentially immobilizing poly(diallyldimethylammonium chloride) and TiO(2) NPs on PDMS surface by layer-by-layer assembly technique. The modified PDMS microchip exhibited a decreased and stable EOF, which was favorable for the separation of biomolecules with similar migration times. Arginine, phenylalanine, serine and threonine were used as model analytes to evaluate the performance of the modified microchip. The four amino acids were efficiently separated within 100 s in a 3.7 cm long separation channel and successfully detected on the carbon fiber electrode in conjunction with in-channel indirect amperometry. Resolutions and theoretical plate numbers of the analytes were considerably enhanced in the presence of TiO(2) NPs. The modified microchip demonstrated excellent stability and reproducibility with improved RSDs of migration times and peak currents for run-to-run, day-to-day and chip-to-chip analyses, respectively. Variables influencing the separation efficiency and amperometric response, including injection and separation voltage, the working electrode position and buffer concentration, were optimized in detail.
Assuntos
Aminoácidos/análise , Dimetilpolisiloxanos/química , Eletroforese em Microchip/instrumentação , Nanopartículas/química , Titânio/química , Eletroforese em Microchip/métodos , Desenho de Equipamento , Limite de Detecção , Reprodutibilidade dos Testes , Propriedades de SuperfícieRESUMO
A novel and simple method based on layer-by-layer (LBL) technique has been developed for the modification of the channel in PDMS electrophoresis microchip to create a hydrophilic surface with a stable EOF. The functional surface was obtained by sequentially immobilizing chitosan and deoxyribonucleic acid (DNA) onto the microfluidic channel surface using the LBL assembly technique. Compared to the native PDMS microchips, the contact angle of the chitosan-DNA modified PDMS microchips decreased and the EOF increased. Experimental conditions were optimized in detail. The chitosan-DNA modified PDMS microchips exhibited good reproducibility and long-term stability. Separation of uric acid (UA) and ascorbic acid (AA) performed on the modified PDMS microchip generated 43,450 and 46,790 N/m theoretical plates compared with 4048 and 19,847 N/m with the native PDMS microchip. In addition, this method has been successfully applied to real human urine samples, without SPE, with recoveries of 97-105% for UA and AA.
Assuntos
Ácido Ascórbico/urina , Dimetilpolisiloxanos/química , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Ácido Úrico/urina , Quitosana/química , DNA/química , Eletroquímica , Humanos , Concentração de Íons de Hidrogênio , Modelos Lineares , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Propriedades de Superfície , Fatores de TempoRESUMO
An effective and facile in situ electroless deposition approach for the fabrication of multi-walled carbon nanotube-supported Prussian blue nanoparticle (MWNTs/PB NP) composite nanomaterials is demonstrated in this article. The coverage of PB NPs on MWNTs is tunable by varying the experimental parameters, such as the initial molar concentration of FeCl3 + K3[Fe(CN)6], the relative concentration of FeCl3 + K3[Fe(CN)6] to MWNTs, and the temperature and duration of the heat treatment. This method involves a simple mixing process followed by a mild heating process and does not need the exhaustive surface oxidation process of MWNTs. TEM, FTIR, UV, and XRD are all used to characterize the MWNTs/PB composite materials. In addition, the electrochemical behavior of PB and catalysis of the reduction of H2O2, are investigated. The novel method is expected to be applicable for preparation of other coordination polymer/MWNTs composites and finds use in applications for electronic nanodevices.
Assuntos
Ferrocianetos/química , Nanocompostos/química , Nanotubos de Carbono/química , Cloretos , Eletrodos , Compostos Férricos/química , Temperatura Alta , Peróxido de Hidrogênio/química , Microscopia Eletrônica de Transmissão , Nanoestruturas/química , Oxigênio/química , Polímeros/química , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura , Difração de Raios XRESUMO
A bimetal lanthanide coordination polymer nanoparticle (ATP-Ce/Tb-Tris CPNs) with good biocompatibility was synthesized in Tris-HCl buffer using adenosine triphosphate (ATP) molecules as the bridge ligands. The large absorption cross section and suitable emission energy of Ce3+ matching to the adsorption energy of Tb3+(4fn) results in the efficient energy transfer from Ce3+ to Tb3+, thus the synthesized ATP-Ce/Tb-Tris CPNs exhibit the characteristic green emission of Tb3+. Such energy transfer from metal to metal in fluorescent lanthanide coordination polymer nanoparticles (Ln-CPNs) has been demonstrated. It is found that the oxidation of Ce3+ in ATP-Ce/Tb-Tris CNPs to Ce4+ would interrupt the energy transfer from Ce3+ to Tb3+, leading to fluorescence quenching of Tb3+. On the basis of this quenching mechanism, ATP-Ce/Tb-Tris CPNs has been successfully used to detect reactive oxygen H2O2 with detection limit as low as 2nM. If glucose oxidase is present in the system, glucose can be determined using the ATP-Ce/Tb-Tris CNPs nanosensor.
Assuntos
Glicemia/análise , Complexos de Coordenação/química , Corantes Fluorescentes/química , Peróxido de Hidrogênio/análise , Elementos da Série dos Lantanídeos/química , Nanopartículas/química , Polímeros/química , Trifosfato de Adenosina/química , Técnicas Biossensoriais/métodos , Cério/química , Transferência de Energia , Glucose Oxidase/química , Humanos , Limite de Detecção , Nanopartículas/ultraestrutura , Espectrometria de Fluorescência/métodos , Térbio/químicaRESUMO
A newly designed molecularly imprinted polymer (MIP) material was developed and successfully used as recognition element for enantioselective recognition by microchip electrophoresis. In this work, molecularly imprinted polymers were facilely prepared employing Fe3O4 nanoparticles (NPs) as the supporting substrate and norepinephrine as the functional monomer in the presence of template molecule in a weak alkaline solution. After extracting the embedded template molecules, the produced imprinted Fe3O4@polynorepinephrine (MIP-Fe3O4@PNE) NPs have cavities complementary to three dimensional shape of template molecules favoring high binding capacity and magnetism property for easy manipulation. The MIP-Fe3O4@PNE NPs prepared with l-tryptophan, l-valine, l-threonine, Gly-l-Phe, S-(-)-ofloxacin or S-(-)-binaphthol as template molecules were packed in the polydimethylsiloxane microchannel via magnetic field as novel stationary phase to successful enantioseparation of corresponding target analysts. The MIP-Fe3O4@PNE NPs-based microchip electrophoresis system exhibited strong recognition ability, excellent high-performance, admirable reproducibility and stability, which provided a powerful protocol for separation enantiomers within a short analytical time and opened up an avenue for multiplex chiral compound assay in various systems.
Assuntos
Nanopartículas de Magnetita/química , Impressão Molecular , Norepinefrina/química , Dimetilpolisiloxanos , Dipeptídeos/química , Eletroforese em Microchip , Ofloxacino/química , Polímeros , Reprodutibilidade dos Testes , Estereoisomerismo , Treonina/química , Triptofano/química , Valina/químicaRESUMO
A novel chip-based enantioselective open-tubular capillary electrochromatography (OT-CEC) was developed employing bovine serum albumin (BSA) conjugated polydopamine-graphene oxide (PDA/GO) nanocomposites (PDA/GO/BSA) as stationary phase. After the poly(dimethylsiloxane) (PDMS) microfluidic chip was filled with a freshly prepared solution containing dopamine and graphene oxide, PDA/GO nanocomposites were formed and deposited on the inner wall of microchannel as permanent coating via the oxidation of dopamine by the oxygen dissolved in the solution. The PDA/GO-coated PDMS microchips not only have the adhesion of PDA that make them easily immobilized in the microchannel, but also have the larger surface and excellent biocompatibility of graphene which can incorporate much more biomolecules and well maintain their biological activity. In addition, incorporation of GO in PDA film can make surface morphology more rough, which is beneficial for enhancing the loading capacity of proteins in the microchannels and increasing sample capacity of OT-CEC columns. BSA was stably immobilized in the PDMS microchannel to fabricate a protein-stationary phase. Compared with the native PDMS microchannels, the modified surfaces exhibited much better wettability, more stable electroosmotic mobility, and less nonspecific adsorption. The efficient separation of chiral amino acids (tryptophan and threonine) and chiral dipeptide demonstrate that the constructed OT-CEC columns own ideal enantioselectivity. The presented strategy using PDA/GO coating as a versatile platform for facile conjugation of proteins may offer new processing strategies to prepare a functional surface designed on microfluidic chips.
Assuntos
Eletrocromatografia Capilar/métodos , Grafite/química , Indóis/química , Óxidos/química , Polímeros/química , Proteínas/química , Proteínas/isolamento & purificação , Adesividade , Adsorção , Dimetilpolisiloxanos/química , Dipeptídeos/química , Dipeptídeos/isolamento & purificação , Eletro-Osmose , Técnicas Analíticas Microfluídicas , Nanocompostos/química , Soroalbumina Bovina/química , Treonina/química , Treonina/isolamento & purificação , Triptofano/química , Triptofano/isolamento & purificaçãoRESUMO
A facile approach for preparation of molecularly imprinted polymers was developed and successfully used as chiral stationary phase for rapid enantioseparation by open tubular capillary electrochromatography (OT-CEC). In this work, molecularly imprinted polymers were one-step prepared employing Fe3O4 nanoparticles (NPs) as the supporting substrate and dopamine as the functional monomer. By simply mixing Fe3O4 NPs with template molecules in a weak alkaline solution of dopamine, a thin adherent polydopamine (PDA) film imprinted with template molecules was formed by the self-polymerization of dopamine on the surface of Fe3O4 NPs. After extracting the embedded template molecules, the produced imprinted Fe3O4@PDA NPs are of three dimensional shape of template molecules favoring high binding capacity and magnetism property for easy manipulation. The imprinted Fe3O4@PDA NPs prepared with l-tryptophan, l-tyrosine, Gly-l-Phe or s-ofloxacin as template molecules were packed in the PDMS microchannel via magnetic field as novel stationary phase for the successful enantioseparation of corresponding target analysts. In addition, the imprinted Fe3O4@PDA NPs-based OT-CEC system exhibited excellent reproducibility, stability and repeatability, which provides a powerful protocol for separation enantiomers within a short analytical time and opens up a promising avenue for high-throughput screening of chiral compounds.
Assuntos
Eletrocromatografia Capilar/métodos , Compostos Férricos/química , Indóis/química , Impressão Molecular/métodos , Nanopartículas/química , Polímeros/química , Fenômenos Magnéticos , Nanopartículas/ultraestrutura , Reprodutibilidade dos Testes , EstereoisomerismoRESUMO
For the first time, a simple and 'green' approach based on one-step strategy was designed and developed for the modification of a fused-silica capillary with polynorepinephrine (PNE) to separate amino acid enantiomers using capillary electrochromatography coupled with electrogenerated chemiluminescence detection (CEC-ECL). Norepinephrine (NE) was filled into capillary to generate PNE coating on the surface of capillary as permanent coating via the oxidation of NE by oxygen dissolvable in the solution. The formation of the PNE coating was characterized by scanning electron microscopy, UV-vis spectra and contact angle measurements. Compared with the native capillary, the modified capillary had much better wettability, less nonspecific adsorption toward amino acids, and the enantiomers of histidine, phenylalanine, and valine samples received baseline separation with the resolution factors of 1.6, 1.8 and 1.6, respectively, utilizing a separation length of 40 cm of the capillary coupled with ECL detection on the PNE-coated capillary.
Assuntos
Eletrocromatografia Capilar/instrumentação , Medições Luminescentes/métodos , Norepinefrina/química , Polímeros/química , Aminoácidos/análise , Aminoácidos/química , Aminoácidos/isolamento & purificação , Eletrocromatografia Capilar/métodos , Limite de Detecção , Norepinefrina/análogos & derivados , Polimerização , Polímeros/síntese química , Reprodutibilidade dos Testes , EstereoisomerismoRESUMO
A novel, simple, and economical method for the preparation of chiral stationary phases for chip-based enantioselective open tubular capillary electrochromatography (OT-CEC) using polydopamine (PDA) coating as an adhesive layer was reported for the first time. After the poly(dimethylsiloxane) (PDMS) microfluidic chip was filled with dopamine (DA) solution, PDA film was gradually formed and deposited on the inner wall of microchannel as permanent coating via the oxidation of DA by the oxygen dissolved in the solution. Due to possessing plentiful catechol and amine functional groups, PDA coating can serve as a versatile multifunctional platform for further secondary reactions, leading to tailoring of the coatings for protein bioconjugation by the thiols and amines via Michael addition or Schiff base reactions. Bovine serum albumin (BSA), acting as a target protein, was then stably and homogeneously immobilized in the PDA-coated PDMS microchannel to fabricate a novel protein stationary phase. Compared with the native PDMS microchannels, the modified surfaces exhibited much better wettability, more stable and enhanced electroosmotic mobility, and less nonspecific adsorption. The water contact angle and electroosmotic flow of PDA/BSA-coated PDMS substrate were measured to be 44° and 2.83×10(-4)cm(2)V(-1)s(-1), compared to those of 112° and 2.10×10(-4)cm(2)V(-1)s(-1) from the untreated one, respectively. Under a mild condition, d- and l-tryptophan were efficiently separated with a resolution of 1.68 within 130s utilizing a separation length of 37mm coupled with in-column amperometric detection on the PDA/BSA-coated PDMS microchips. This present versatile platform, facile conjugation of biomolecules onto microchip surfaces via mussel adhesive protein inspired coatings, may offer new processing strategies to prepare a biomimetic surface design on microfluidic chips, which is promising in high-throughput and complex biological analysis.
Assuntos
Eletrocromatografia Capilar/métodos , Indóis/química , Polímeros/química , Soroalbumina Bovina/isolamento & purificação , Adsorção , Animais , Eletrocromatografia Capilar/instrumentação , Bovinos , Soroalbumina Bovina/química , EstereoisomerismoRESUMO
In this study, a novel biomolecule immobilization approach has been proposed to the synthesis of multi-functional core-shell glucose oxidase-Au-polydopamine-Fe3O4 magnetic bionanoparticles (GOx-Au-PDA-Fe3O4 MBNPs) using the one-pot chemical polymerization method. Then, a high performance biosensor has been constructed by effectively attaching the proposed GOx-Au-PDA-Fe3O4 MBNPs to the surface of the magnetic glassy carbon electrode. Scanning electron microscope, energy dispersive x-ray spectrometer, UV-vis spectroscopy, and electrochemical methods were used to characterize the GOx-Au-PDA-Fe3O4 MBNPs. The resultant GOx-Au-PDA-Fe3O4 MBNPs not only have the magnetism of Fe3O4 nanoparticles which makes them easily manipulated by an external magnetic field, but also have the excellent biocompatibility of PDA to maintain the native structure of the GOx, and good conductivity of Au nanoparticles which can facilitate the direct electrochemistry of GOx in the biofilm. Hence, the present GOx-Au-PDA-Fe3O4 biofilm displays good linear amperometric response to glucose concentration ranging from 0.02 to 1.875 mM. This efficient biomolecule immobilization platform is recommended for the preparation of many other MBNPs with interesting properties and application potentials in many fields, such as biosensing, biocatalysis, biofuel cells, and bioaffinity separation.
Assuntos
Técnicas Biossensoriais/métodos , Glucose Oxidase/química , Glucose/isolamento & purificação , Nanopartículas de Magnetita/química , Quitosana/química , Enzimas Imobilizadas , Compostos Férricos/química , Ouro/química , Indóis/química , Polímeros/químicaRESUMO
In this work, the magnetic core-shell Fe(3)O(4)@Au nanoparticles attached to the surface of a magnetic glassy carbon electrode (MGCE) were applied to the immobilization/adsorption of myoglobin (Mb) for fabricating Mb/Fe(3)O(4)@Au biofilm. The morphology, structure, and electrochemistry of the nanocomposite were characterized by transmission electron microscope, UV-vis spectroscopy, electrochemical impedance spectroscopy, and cyclic voltammetry, respectively. The resultant Fe(3)O(4)@Au NPs not only have the magnetism of Fe(3)O(4) NPs that make them easily manipulated by an external magnetic field, but also have the good conductivity and excellent biocompatibility of Au layer which can maintain the bioactivity and facilitate the direct electrochemistry of Mb in the biofilm. The modified electrode based on this Mb/Fe(3)O(4)@Au biofilm displayed good electrocatalytic activity to the reduction of H(2)O(2) with a linear range from 1.28 to 283 microM. The proposed method simplified the immobilization methodology of proteins and showed potential application for fabricating novel biosensors and bioelectronic devices.