Quartz crystal microbalance immunosensor for the detection of antibodies to double-stranded DNA

We report the development of a novel quartz crystal microbalance immunosensor with the simultaneous measurement of resonance frequency and motional resistance for the detection of antibodies to double-stranded DNA (dsDNA). The immobilization of poly(l-lysine) and subsequent complexation with DNA resulted in formation of a sensitive dsDNA-containing nanofilm on the surface of a gold electrode. Atomic force microscopy has been applied for the characterization of a poly(l-lysine)–DNA film. After the blocking with bovine serum albumin, the immunosensor in flow-injection mode was used to detect the antibodies to dsDNA in purified protein solutions of antibodies to dsDNA and to single-stranded DNA, monoclonal human immunoglobulin G, DNase I and in blood serum of patients with bronchial asthma and systemic lupus erythematosus. Experimental results indicate high sensitivity and selectivity of the immunosensor. In memoriam Prof. Victor G. Vinter     

Nuclear magnetic resonance of mass-limited samples using small RF coils

Figure Schematic diagram of a typical arrangement used for hyphenating chemical microseparations (e.g. capillary HPLC, CE, or CEC) with microcoil NMR detection     

Simultaneous determination of uric acid and ascorbic acid at a ferrocenium–thioglycollate modified electrode

Self-assembled monolayers (SAMS) of chemisorbed thioglycollate on a gold electrode surface have been used as a base interface for the electrostatic adsorption of ferrocenium ion. Electrochemical impedance spectra (EIS) and cyclic voltammetry (CV) were used to evaluate the electrochemical properties of the supramolecular film. The bare gold electrode failed to distinguish the oxidation peaks of ascorbic acid (AA) and uric acid (UA) in phosphate buffer solution (PBS, pH 7.0), while the ferricinium–thioglycollate modified electrode could separate them efficiently. In differiential pulse voltammetric measurements, the prepared gold electrode could separate AA and UA signals, allowing the simultaneous determination of AA and UA. Under optimal conditions and within the linear range of 1.0 × 10⁻⁶ to 5.0 × 10⁻⁴ M, the detection limits of AA and UA achieved were 2.0 × 10⁻⁷ and 1.0 × 10⁻⁷ M, respectively. The applicability of the prepared electrode was demonstrated by measuring AA and UA in human urine without any pretreatment. Figure Fabrication process for the modified electrode     

Differential pulse voltammetric determination of dopamine with the coexistence of ascorbic acid on boron-doped diamond surface

Electrochemical determination of dopamine (DA) in the presence of ascorbic acid (AA) was achieved on boron-doped diamond (BDD) film electrode by differential pulse voltammetry. The experimental results indicated that the oxidative peaks of DA and AA could be separated completely on anodically-treated (BDD) electrode without further modification, although these two peaks can not be separated on glassy carbon electrode. The peak separation of DA and AA was developed to be 0.44 V. High sensitivity was obtained to determine DA selectively with the coexisting of a large excess of AA in acidic media by DPV. The detection limit of DA was achieved to be 1.1 × 10^-6 M in the presence of AA with the concentration of 200 times more than DA. This technique was also applied to the determination of DA in real samples.      

Simultaneous determination of ascorbic acid,dopamine, and uric acid using a carbon paste electrode modified with multiwalled carbon nanotubes,ionic liquid,and palladium nanoparticles

We describe the modification of a carbon paste electrode (CPE) with multiwalled carbon nanotubes (MWCNT) and an ionic liquid (IL). Electrochemical studies revealed an optimized composition of 60 % graphite, 20 % paraffin, 10 % MWCNT and 10 % IL. In a next step, the optimized CPE was modified with palladium nanoparticles (Pd-NPs) by applying a double-pulse electrochemical technique. The resulting electrode was characterized by scanning electron microscopy, energy dispersive X-ray spectroscopy, X-ray diffraction, cyclic voltammetry, and electrochemical impedance spectroscopy. It gives three sharp and well separated oxidation peaks for ascorbic acid (AA), dopamine (DA), and uric acid (UA), with peak separations of 180 and 200 mV for AA-DA and DA-UA, respectively. The sensor enables simultaneous determination of AA, DA and UA with linear responses from 0.6 to 112, 0.1 to 151, and 0.5 to 225 μM, respectively, and with 200, 30 and 150 nM detection limits (at an S/N of 3). The method was successfully applied to the determination of AA, DA, and UA in spiked samples of human serum and urine.

The CPE was modified with multiwalled carbon nanotubes and an ionic liquid. After optimization the electrode was further modified with palladium nanoparticles. The resulting electrode gives three sharp and well separated oxidation peaks for ascorbic acid, dopamine and uric acid

     

Ultra-sensitive voltammetric simultaneous determination of dopamine,uric acid and ascorbic acid based on a graphene-coated alumina electrode

The authors describe the fabrication of an interconnected edge-exposed graphene nanostructure via chemical vapor deposition (CVD) of foliated graphene onto a network of alumina nanofibers. The fibers such obtained are shown to enable ultra-sensitive voltammetric determination of dopamine (DA), uric acid (UA) and ascorbic acid (AA). The electrode displays powerful and persistent electro oxidative behavior and excellent electron transport properties. Cyclic voltammetry and differential pulse voltammetry demonstrate excellent selectively and sensitivity for AA, DA and UA, with typical peaks at ?0.08 V, +0.19 V, and +0.34 V (vs. SCE), respectively. Under optimum conditions, the calibration plots are linear in the 1–80 μM range for DA, in the 1–60 μM range for UA, and in the 0.5–60 μM range for UA, with detection limits of 0.47 μM, 0.28 μM and 0.59 μM, respectively. The sensor was successfully applied to the simultaneous determination of DA and UA in the presence of AA in spiked urine sample.

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Graphical abstract Material with high density of graphene foliates grown over highly aligned nano-dimensional ceramic fibers is used as electrode for simultaneous highly sensitive electrochemical determination of DA in the presence of UA and AA with a considerably low limit of detection.

     

Polycation coating poly(dimethylsiloxane) capillary electrophoresis microchip for rapid separation of ascorbic acid and uric acid

A novel method for rapid separation and determination of ascorbic acid and uric acid has been developed with a polycation-modified poly(dimethylsiloxane) (PDMS) microchip under a negative-separation electric field. Just by flushing the microchip with aqueous solutions of the polycations, poly(allylamine) hydrochloride, poly(diallyldimethylammonium chloride) or chitosan could be stably coated on the PDMS microchannel surface, which resulted in a reversed electroosmotic flow and thus the rapid and efficient separation of the two substrates. Factors influencing the separation, including polycation category, buffer solution, detection potential and separation voltage, were investigated and optimized. The cheapness, rapid analysis speed and the successful analysis of human urine make this microsystem attractive for application in clinics. Figure The electropherograms of 100 μ/mL AA and UA in (1) PAH, (2) PDDA, (3) Chitosan modified PDMS microchannels and native PDMS microchip (4).     

Bienzyme sensors based on novel polymethylferrocenyl dendrimers

Amperometric bienzyme electrodes with horseradish peroxidase (HRP) and glucose oxidase (GOx) co-immobilized on polymethylferrocenyl dendrimers deposited onto platinum electrodes have been used for determination of the hydrogen peroxide produced by the oxidase during the enzymatic reaction. The redox dendrimers consist of flexible poly(propylenimine) dendrimer cores functionalised with octamethylferrocenyl units. The effects of dendrimer generation, the thickness of the dendrimer layer, substrate concentration, interferences, and reproducibility on the response of the sensors were investigated. The new bienzyme biosensors respond to substrate at work potential values between 200 and 50 mV (vs. SCE), have good sensitivity, and are resistant to interferences. Figure     

Simultaneous use of multiple fluorescent reporter dyes for glucose sensing in aqueous solution

The simultaneous use of several fluorescent reporter dyes in a multicomponent boronic acid-based glucose sensing system is reported. In one application, two dyes with widely different emission wavelengths are used to report changes in glucose concentration. A third glucose-insensitive dye was then added to act as a reference dye and provide for a ratiometric correction to the two reporter dye signals. The inclusion of such a reference dye reduces errors arising from sources such as fluctuations in lamp intensity and sample dilution. The simultaneous use of multiple fluorescent reporter dyes     

A glassy carbon electrode modified with MoS<Subscript>2</Subscript> nanosheets and poly(3,4-ethylenedioxythiophene) for simultaneous electrochemical detection of ascorbic acid,dopamine and uric acid

We describe a chemical exfoliation method for the preparation of MoS₂ nanosheets. The nanosheets were incorporated into poly(3,4-ethylenedioxythiophene) (PEDOT) by electrodeposition on a glassy carbon electrode (GCE) to form a nanocomposite. The modified GCE is shown to enable simultaneous determination of ascorbic acid (AA), dopamine (DA) and uric acid (UA). Due to the synergistic effect of MoS₂ and PEDOT, this electrode displays better properties in terms of electrocatalytic oxidation of AA, DA and UA than pure PEDOT, which is illustrated by cyclic voltammetry and differential pulse voltammetry (DPV). Under optimum conditions and at pH 7.4, the respective sensitivities and best working potentials are as follows: AA: 1.20 A?mM^?1?m^?2, 30 mV; DA: 36.40 A?mM^?1?m^?2, 210 mV; UA: 105.17 A?mM^?1?m^?2, 350 mV. The calculated detection limits for AA, DA and UA are 5.83 μM, 0.52 μM and 0.95 μM, respectively. The modified electrode was applied to the detection of the three species in human urine samples and gave satisfactory results.

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Graphical abstract MoS₂ nanosheets were prepared by a facile chemical exfoliation method. MoS₂ and poly(3,4-ethylenedioxythiophene) nanocomposite modified glassy carbon electrodes were fabricated, which are shown to enable simultaneous determination of ascorbic acid, dopamine and uric acid with high sensitivity and selectivity.

     

Rapid and simple electrochemical detection of morphine on graphene–palladium-hybrid-modified glassy carbon electrode

A hybrid of reduced graphene oxide–palladium (RGO–Pd) nano- to submicron-scale particles was simultaneously chemically prepared using microwave irradiation. The electrochemical investigation of the resulting hybrid was achieved using cyclic voltammetry and differential pulse voltammetry. RGO–Pd had a higher current response than unmodified RGO toward the oxidation of morphine. Several factors that can affect the electrochemical response were studied, including accumulation time and potential, Pd loading, scan rate, and pH of electrolyte. At the optimum conditions, the concentration of morphine was determined using differential pulse voltammetry in a linear range from 0.34 to 12 μmol L^?1 and from 14 to 100 μmol L^?1, with detection limits of 12.95 nmol L^?1 for the first range. The electrode had high sensitivity toward morphine oxidation in the presence of dopamine (DA) and of the interference compounds ascorbic acid (AA) and uric acid (UA). Electrochemical determination of morphine in a spiked urine sample was performed, and a low detection limit was obtained. Validation conditions including reproducibility, sensitivity, and recovery were evaluated successfully in the determination of morphine in diluted human urine.

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Electrochemical preparation of copper–dendrimer nanocomposites: picomolar detection of Cu<Superscript>2+</Superscript> ions

The present work describes, for the first time, in situ electrochemical preparation of dendrimer-encapsulated Cu nanoparticles using a self-assembled monolayer of fourth-generation amine-terminated polyamidoamine (PAMAM) dendrimer as the template. Atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) studies of the modified surface confirmed the presence of Cu nanoparticles entrapped in dendrimer film. Au electrode modified with a monolayer of the dendrimer enables preconcentration and subsequent voltammetric detection of Cu²⁺ at picomolar concentrations. Further, Cu nanoparticles in the dendrimer monolayer could be electrochemically derivatised to Cu hexacyanoferrate, which exhibits specific crystal planes, unlike the random distribution of crystal planes in bulk-formed Cu hexacyanoferrate, which is another catalytically active material for sensor applications. Figure Electrochemical preparation of copper–dendrimer nanocomposite     

Evaluation of quantum dots applied as switchable layer in a light-controlled electrochemical sensor

Gold electrodes with switchable conductance are created by coating the gold surface with different colloidal quantum dots. For the quantum dot immobilization, a dithiol compound was used. By polarizing the electrode and applying a light pointer, local photocurrents were generated. The performance of this setup was characterized for a variety of different nanoparticle materials regarding drift and signal-to-noise ratio. We varied the following parameters: quantum dot materials and immobilization protocol. The results indicate that the performance of the sensor strongly depends on how the quantum dots are bound to the gold electrode. The best results were obtained by inclusion of an additional polyelectrolyte film, which had been fabricated using layer-by-layer assembly.      

On-line NMR detection of microgram quantities of heparin-derived oligosaccharides and their structure elucidation by microcoil NMR

The isolation and purification of sufficient quantities of heparin-derived oligosaccharides for characterization by NMR is a tedious and time-consuming process. In addition, the structural complexity and microheterogeneity of heparin makes its characterization a challenging task. The improved mass-sensitivity of microcoil NMR probe technology makes this technique well suited for characterization of mass-limited heparin-derived oligosaccharides. Although microcoil probes have poorer concentration sensitivity than conventional NMR probes, this limitation can be overcome by coupling capillary isotachophoresis (cITP) with on-line microcoil NMR detection (cITP-NMR). Strategies to improve the sensitivity of on-line NMR detection through changes in probe design and in the cITP-NMR experimental protocol are discussed. These improvements in sensitivity allow acquisition of cITP-NMR survey spectra facilitating tentative identification of unknown oligosaccharides. Complete structure elucidation for microgram quantities of the purified material can be carried out through acquisition of 2D NMR spectra using a CapNMR microcoil probe. Survey NMR spectrum obtained by cITP-NMR using a second-generation probe (the microcoil of which is shown) facilitates tentative identification of unknown oligosaccharides (e.g., the heparin-derived tetrasaccharide illustrated)     

H-point standard addition method applied to solid-state stripping voltammetry: quantitation of lead and tin in archaeological glazes

A solid-state electrochemical application of the H-point standard addition method to the quantification of two depositable metals A and B, which produce strongly overlapped stripping peaks, is described. The method is based on the mechanical transference of mixtures of the solid sample plus a selected compound, of a reference depositable metal R, and of known amounts of a reference material containing A or B, to paraffin-impregnated graphite electrodes. After a reductive deposition step, voltammograms recorded for those modified electrodes immersed into a suitable electrolyte produce stripping peaks for the oxidation of all of the metals deposited. Measurement of the currents at selected potentials in overlapping peaks corresponding to the stripping of A and B permits the quantitation of these metals in the solid sample, while avoiding matrix effects. The method was applied to the simultaneous determination of Pb and Sn in archaeological glazes using PbCO₃ and SnO₂ as standards and ZnO as a reference material.      

Detection of flavonoids and assay for their antioxidant activity based on enlargement of gold nanoparticles

We report our findings that natural flavonoids such as quercetin, daizeol and puerarin can act as reductants for the enlargement of gold nanoparticles (Au-NPs). Consequently, the UV–vis spectra of a solution containing Au-NPs will be gradually changed, and the molecules of the natural herbs can be detected by making use of changes in the UV–visible spectra. Furthermore, we have prepared a self-assembled monolayer modified electrode by modifying cysteamine on a gold substrate electrode, which is further modified by some Au-NP seeds. When the modified electrode is immersed in a solution containing flavonoids and tetrachloroauric acid as a gold source for the growth of the Au-NP seeds, with the increase of the concentration of flavonoids, the Au-NP seeds on the surface of the modified electrode can be enlarged to varying degrees. As a result, the peak currents in the corresponding cyclic voltammograms are inversely decreased, and simultaneously the peak separation is increased. Therefore, an electrochemical method to detect flavonoids is also proposed. Compared with the optical detection method, the electrochemical method has an extraordinarily lower detection limit and a significantly extended detection range. Moreover, the optical and electrochemical experimental results can be also used to assay and compare the relative antioxidant activities of the flavonoids. Figure Enlargement of Au nanoparticles by flavonoids at cysteamine modified electrode     

Evaporative light scattering detection: trends in its analytical uses

Evaporative light scattering detection (ELSD) is widely recognized as a universal tool for liquid and supercritical chromatographies. In addition, this detection technique is fully compatible with continuous-flow systems. In fact, the combination of continuous non-chromatographic techniques and ELSD affords the design of simple, reliable systems for extracting qualitative information. This paper reviews instrumental innovations regarding the miniaturization of evaporative light scattering detectors and their uses in micro and capillary liquid chromatography; also, it discusses their increasingly important role in the development of vanguard configurations for sample screening and the determination of total indices without the need for chromatographic separation. Moreover, it compares them with other types of chromatographic detectors in terms of performance. Finally, the potential of ELSD for solving real-life analytical problems arising from the need to meet (bio)chemical information needs is illustrated with various selected applications. Figure New trends in evaporative light scattering detection: recent chromatographic uses and its role in vanguard/rearguard strategies     

Determination of acetone in seawater using derivatization solid-phase microextraction

Acetone plays an important role in the chemistry of both the atmosphere and the ocean, due to its potential effect on the tropospheric HO_x (= HO + HO₂) budget, as well as its environmental and health effects. We discuss the development of a mobile, sensitive, selective, economical and facile method for the determination of acetone in seawater. The method consists of derivatizing acetone to its pentafluorobenzyl oxime using 1,2,3,4,5-pentafluorobenzylhydroxylamine (PFBHA), followed by solid-phase microextraction (SPME) and analysis by gas chromatography/mass spectrometry (GC/MS). A detection limit of 3.0 nM was achieved. The buffering capacity of seawater imposes challenges in using the method’s optimum pH (3.7) on seawater samples, requiring calibration standards to be made in buffered salt water and the acidification of seawater samples and standards prior to extraction. We employed the technique for analysis of selected surface seawater samples taken on the Nordic seas during the ARK-XX/1 cruise (R.V. Polarstern). An upper limit of 5.5–9.6 nM was observed for acetone in these waters, the first acetone measurements reported for far North Atlantic and Arctic waters. Simplified schematic of transformations of organic compounds at the atmosphere–ocean interface     

Simultaneous determination of dopamine and uric acid in the presence of ascorbic acid using a gold electrode modified with carboxylated graphene and silver nanocube functionalized polydopamine nanospheres

A voltammetric sensor is presented for the simultaneous determination of dopamine (DA) and uric acid (UA) in the presence of ascorbic acid (AA). It is based on a gold electrode (GE) modified with carboxyl-functionalized graphene (CFG) and silver nanocube functionalized DA nanospheres (AgNC@PDA-NS). The AgNC@PDA-NS nanocomposite was characterized by scanning electron microscopy and UV-Vis spectroscopy. The electrochemical behavior of the modified electrode was evaluated by electrochemical impedance spectroscopy, cyclic voltammetry and differential pulse voltammetry. The modified electrode displays good electrocatalytic activity towards DA (typically at 0.14 V vs. Ag/AgCl) and UA (typically at 0.29 V vs. Ag/AgCl) even in the presence of ascorbic acid. Response to DA is linear in the concentration range of 2.5 to 130 μM with a detection limit of 0.25 μM. Response to UA is linear in the concentration range of 10 to 130 μM with a detection limit of 1.9 μM. In addition, the sensitivity for DA and UA is 0.538 and 0.156 μA μM^?1 cm^?2, respectively. The modified electrode also displays good stability, selectivity and reproducibility.

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Graphical abstract The gold electrode modified with polydopamine nanospheres functionalized with silver nanocube and carboxylated graphene is used for simultaneous determination of DA and UA in the presence of AA, with wide linear range and low detection limit.

     

A low perfusion rate microreactor for continuous monitoring of enzyme characteristics: application to glucose oxidase

This report describes a versatile and robust microreactor for bioactive proteins physically immobilized on a polyether sulfone filter. The potential of the reactor is illustrated with glucose oxidase immobilized on a filter with a cut-off value of 30 kDa. A flow-injection system was used to deliver the reactants and the device was linked on-line to an electrochemical detector. The microreactor was used for on-line preparation of apoglucose oxidase in strong acid and its subsequent reactivation with flavin adenine dinucleotide. In addition we describe a miniaturized version of the microreactor used to assess several characteristics of femtomole to attomole amounts of glucose oxidase. A low negative potential over the electrodes was used when ferrocene was the mediator in combination with horseradish peroxidase, ensuring the absence of oxidation of electro-active compounds in biological fluids. A low backpressure at very low flow rates is an advantage, which increases the sensitivity. A variety of further applications of the microreactor are suggested. Figure Preparation of apoGOx and restoration of enzyme activity using a soluton of FAD