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An amperometric detector formed of highly dispersed Ni nanoparticles embedded in a graphite-like carbon film electrode for sugar determination.

We achieved improved detection limits for sugars by developing a novel thin film containing 0.8% highly dispersed Ni nanoparticles in disordered graphite-like carbon (Ni-NDC) as a detection electrode for high-performance liquid chromatography. The Ni-NDC film was prepared in one step by a simple radio frequency (rf) sputtering method at a temperature below 200 degrees C. We characterized the film by XPS, TEM, and AFM analysis and found that the average Ni nanoparticle size was 3 nm and that the film consisted of a mixture of Ni, NiO, Ni2O3, and Ni(OH)2. We studied the electrochemical detection of sugars using the 0.8% Ni-NDC film electrode. The film electrode had excellent electrocatalytic ability and good stability compared with a Ni-bulk electrode with regard to the electrooxidation of sugars. We employed the Ni-NDC film as an HPLC detection electrode. We achieved a good separation of four sugars (glucose, fructose, sucrose, lactose) at a relatively low constant detection potential (0.40 V vs Ag/AgCl) and a linearity of over 3 orders of magnitude. We obtained improved detection limits for the investigated sugars, namely, 20, 25, 50, and 37 nM for glucose, fructose, sucrose, and lactose, respectively. This is at least 1 order of magnitude lower than the detection limits obtained with a Ni-bulk electrode with the same measurement condition. The Ni-NDC film electrode also showed good reproducibility with a relative standard deviation of 1.75% for 40 consecutive injections of glucose in a flow system.

Carbon↗

Liquid / liquid ion-transfer processes at the dioctylphosphoric acid (N,N-didodecyl-N',N'-diethylphenylenediamine) / water (electrolyte) interface at graphite and mesoporous TiO2 substrates.

Biphasic electrode systems are studied for the case of the oxidation of the water-insoluble liquid N,N-didodecyl-N',N'-diethylphenylenediamine (DDPD) neat and dissolved in bis(2-ethylhexyl) phosphate (HDOP) and immersed in aqueous electrolyte media. The oxidation process in the absence of HDOP is accompanied by transfer of the anion (perchlorate or phosphate) from the water into the organic phase. However, in the presence of HDOP, oxidation is accompanied by proton exchange instead. This electrochemically driven proton exchange process occurs over a wide pH range. Organic microdroplet deposits of DDPD in HDOP at basal plane pyrolytic graphite electrodes are studied by voltammetric techniques and compared in their behavior to organic microphase deposits in mesoporous TiO2 thin films. The mesoporous TiO2 thin film acts as a host for the organic liquid and provides an alternative biphasic electrode system compared to the random microdroplet/graphite system. Two types of mesoporous TiO2 thin-film electrodes, (i) a 300-400-nm film on ITO and (ii) a 300-400-nm film on ITO sputter-coated with a 20-nm porous gold layer, are investigated.

Electrochemistry↗

Electrochemical reduction of NO by hemin adsorbed at pyrolitic graphite.

The mechanism of the electrochemical reduction of nitric oxide (NO) by hemin adsorbed at pyrolitic graphite was investigated. The selectivity of NO reduction was probed by combining the rotating ring disk electrode (RRDE) technique with a newly developed technique called on-line electrochemical mass spectroscopy (OLEMS). These techniques show that NO reduction by adsorbed heme groups results in production of hydroxylamine (NH(2)OH) with almost 100% selectivity at low potentials. Small amounts of nitrous oxide (N(2)O) were only observed at higher potentials. The rate-determining step in NO reduction most likely consists of an electrochemical equilibrium involving a proton transfer, as can be derived from the Tafel slope value of 62 mV/dec and the pH dependence of -42 mV/pH. The almost 100% selectivity toward NH(2)OH distinguishes this system both from NO reduction on bare metal electrodes, which often yields NH(3), and from biological NO reduction in cytochrome P450nor, which yields N(2)O exclusively.

Adsorption↗

Electronic and geometric properties of Au nanoparticles on Highly Ordered Pyrolytic Graphite (HOPG) studied using X-ray Photoelectron Spectroscopy (XPS) and Scanning Tunneling Microscopy (STM).

Au nanoparticles grown on mildly sputtered Highly Ordered Pyrolytic Graphite (HOPG) surfaces were studied using Scanning Tunneling Microscopy (STM) and X-ray Photoelectron Spectroscopy (XPS). The results were compared with those of Ag nanoparticles on the same substrate. By varying the defect densities of HOPG and the Au coverages, one can create Au nanoparticles in various sizes. At high Au coverages, the structures of the Au films significantly deviate from the ideal truncated octahedral form: the existence of many steps between different Au atomic layers can be observed, most likely due to a high activation barrier of the diffusion of Au atoms across the step edges. This implies that the particle growth at room temperature is strongly limited by kinetic factors. Hexagonal shapes of Au structures could be identified, indicating preferential growth of Au nanostructures along the (111) direction normal to the surface. In the case of Au, XPS studies reveal a weaker core level shift with decreasing particle size compared to the 3d level in similarly sized Ag particles. Also taking into account the Auger analysis of the Ag particles, the core level shifts of the metal nanoparticles on HOPG can be understood in terms of the metal/substrate charge transfer. Ag is (partially) positively charged, whereas Au negatively charged on HOPG. It is demonstrated that XPS can be a useful tool to study metal-support interactions, which plays an important role for heterogeneous catalysis, for example.

Electrochemistry↗

Chain-length effects on molecular conformation in and chirality of self-assembled monolayers of alkoxylated benzo[c]cinnoline derivatives on highly oriented pyrolytic graphite.

Self-assembled structures of alkoxylated benzo[c]cinnoline derivatives prepared on highly oriented pyrolytic graphite at room temperature from their solutions in solvents such as 1-phenyloctane, toluene, and 1-octanol were studied by scanning tunneling microscopy. The alkoxy chain length markedly affected the molecular conformations in 2-dimensional assemblies of these derivatives. Long-chain derivatives adopted the trans conformations more often than cis, whereas short-chain derivatives took exclusively the cis conformations in the self-assembled monolayers (SAMs). For the derivatives of intermediate chain lengths, polymorphism existed, with four molecular conformations identified experimentally. Experimental evidence substantiated the formation of chiral SAM structures at the surface, which can be explained by the conformations of the molecules. The chirality was also affected by the chain length of the molecules. A simple method analyzing the angles between different domains in the SAMs was used to identify the molecular conformations and to predict their relative structures.

Graphite↗

Molecular dynamics simulation of the adsorption of a fibronectin module on a graphite surface.

We report atomistic simulations of the adsorption of a fibronectin type I module on a hydrophobic graphite surface. This module comprises only beta-sheets, unlike the albumin fragments previously investigated by us which contained only alpha-helices (Raffaini, G.; Ganazzoli, F. Langmuir 2003, 19, 3403-3412). As done in the latter case, most simulations are carried out in an effective dielectric medium by energy minimizations and molecular dynamics (MD). Further optimizations and MD runs in the explicit presence of water are also performed to assess the stability of the geometries found and to describe the solvation of the adsorbed fibronectin module. The initial adsorption is accompanied by local rearrangements of the strands in contact with the surface, but the overall molecular structure is largely preserved. Much larger rearrangements take place at longer times as found through the MD runs, with the molecule spreading as much as possible so as to maximize the surface coverage, hence the interaction energy, despite a significant strain energy. Energetic aspects of adsorption together with the concomitant size change are discussed in comparison with our previous results for two albumin fragments.

Adsorption↗

Porphyrinphosphonate fibers on mica and molecular rows on graphite.

meso-Tetra(phenyl-p-phosphonate) porphyrin forms rigid and well-separated fibers of monomolecular thickness (2.8 nm) and lengths of several micrometers on mica at pH 13 (octasodium salt). The formation of these fibers could be observed directly by tapping mode scanning force microscopy (SFM) and was induced by capillary forces. Normal height images or images with a topographical inversion were observed depending on the distance of the SFM tip. Amplitude-distance curves indicated that a stable meniscus was formed on hydrophilic surface areas below a tip-sample separation of 20 nm. The meniscus let the original nanorods appear as ditches in the mica surface and enabled rearrangements. A partly protonated form of the same porphyrin (pH 11.5) gave rows of flat-lying porphyrins on graphite, which appear with molecular resolution in SFM images as well as two-dimensional platelets of monomolecular thickness.

Aluminum Silicates↗

Electrochemistry using self-assembled DNA monolayers on highly oriented pyrolytic graphite.

Duplex DNA functionalized with pyrene has been utilized to fabricate DNA-modified electrodes on highly oriented pyrolytic graphite (HOPG). Films have been characterized using AFM and radioactive labeling as well as electrochemically. The data obtained are consistent with a close-packed structure in the film with helices oriented in a nearly upright orientation, as seen earlier with the fabrication of thiol-tethered duplexes on gold. Also as on gold, we observe the reduction of DNA-bound intercalators in a DNA-mediated reaction. The reduction of the intercalator is attenuated in the presence of the single-base mismatches, CA and GT, independent of the sequence composition of the oligonucleotide. This sensitivity to single-base mismatches is enhanced when methylene blue reduction is coupled in an electrocatalytic cycle with ferricyanide. The extended potential range afforded by the HOPG surface has allowed us also to investigate the electrochemistry of previously inaccessible metallointercalators, Ru(bpy)2dppz2+ and Os(phen)2dppz2+, at the DNA-modified HOPG surface. These results support the application of DNA-modified HOPG as a convenient and reproducible surface for electrochemical DNA sensors using DNA-mediated charge transport.

Cross-Linking Reagents↗

Coadsorption of guanine and cytosine on graphite: ordered structure based on GC pairing.

Nanostructures formed by coadsorption of the complementary DNA bases guanine (G) and cytosine (C) at a graphite surface in 1-octanol solvent were investigated by in situ scanning tunneling microscopy. The high-resolution observations showed for the first time a well-ordered coadsorption structure, attributed to rows formed from Watson-Crick G-C pairs, which was distinctly different from the structures observed for the individual G/C components. The observed coadsorption structure has been modeled by self-consistent charge density-functional-based tight-binding (SCC-DFTB) calculations, providing information on the intermolecular interactions underlying its formation.

Adsorption↗

Hydrogen sieving and storage in fullerene intercalated graphite.

The geometrical properties of recently synthesised C60 intercalated in graphite have been confirmed by density-functional-based computer simulations. The capability of this material to store molecular hydrogen by physisorption is evaluated. While the material can sieve H2 from heavier molecular gases, our free energy calculations indicate that further tuning of the system by reducing the amount of intercalated fullerene cages is necessary to achieve H2 loadings which are interesting for technical applications.

Fullerenes↗

Self-gelatinizable copolymer immobilized glucose biosensor based on prussian blue modified graphite electrode.

A novel poly(vinyl alcohol) grafting 4-vinylpyridine self-gelatinizable copolymer was adapted to immobilize glucose oxidase. The reduction of hydrogen peroxide (H2O2) was detected at a Prussian Blue (PB) modified graphite electrode. A stable and sensitive glucose amperometric biosensor is described. The copolymer is a good biocompatible polymer in which the glucose oxidase retains high activity. Moreover, the copolymer can adhere firmly to the inorganic PB membrane. The sensor showed an apparent Michaelis-Menten constant of 18 +/- 0.2 mM and a maximum current density of 1.14 microA cm-2 mM-1. The linear range is from 5 microM to 4.5 mM glucose and the detection limit is 0.5 microM glucose. The catalytic efficiency of PB for the reduction of H2O2 is higher than that for the oxidation of H2O2. Glucose concentrations in serum samples from healthy persons and diabetic patients were determined using the sensor. The results compared well with those provided by the hospital using a spectroscopy method.

Biosensing Techniques↗

[Os(bpy)2(PVP)10Cl]Cl polymer and Nafion dual-film modified graphite electrode for the amperometric determination of trace amounts of norepinephrine.

A chemically modified graphite electrode was prepared by using a dual film of [Os(bpy)2(PVP)10Cl]Cl polymer and Nafion. The modified electrode showed excellent electrocatalytical activity for the oxidation of norepinephrine (NE) and an ability to eliminate efficiently the interference of ascorbic acid and other anions. The catalytic peak currents obtained from the cyclic voltammograms increased linearly with increasing concentration of NE. A log-log plot of catalytic current versus NE concentration showed a dual-linear relationship in the ranges 1.8 x 10(-8)-4.4 x 10(-6) M and 4.4 x 10(-6)-2.9 x 10(-4) M with correlation coefficients of 0.990 and 0.999, respectively. The detection limit was about 18 nM (3 delta). At a potential of +500 mV the chronoamperometric response showed a linear relationship between the steady state current and NE concentration in the range 1.3-130 microM. With a further increase in NE concentration a Michaelis-Menten-shaped response was observed. The apparent Michaelis-Menten constant and the maximum current were 1.7 mM and 86 microA, respectively. The modified electrode showed excellent reproducibility, sensitivity and stability for the determination of NE at trace levels.

Electrochemistry↗

Electrocatalytic detection of streptomycin and related antibiotics at ruthenium dioxide modified graphite--epoxy composite electrodes.

The application of ruthenium dioxide (RuO2) modified electrodes to the electrocatalytic detection of the saccharide-related antibiotics streptomycin, novobiocin and neomycin, at low fixed potentials, was investigated. The RuO2-modified graphite - epoxy composite electrodes give extremely stable and reproducible catalytic oxidation currents for these antibiotics at potentials as low as +0.2 V (versus Ag - AgCl). Rapid quantification at the micromolar level is therefore possible. Standard calibration graphs for streptomycin and neomycin yielded slopes of 4.43 and 0.08 nA microM-1 over the linear ranges of 1.5 x 10(-6) - 2.5 x 10(-4) and 1 x 10(-5) - 2 x 10(-3) M, respectively. Owing to its catalytic oxidation by the RuIII - RuIV couple, rather than the RuIV - RuVI transition (which catalyses the oxidation of streptomycin and neomycin), novobiocin could be detected at a lower (+0.2 V) potential, with a sensitivity of 1.31 nA microM-1. Detection limits of 1.5, 6.0 and 10 microM were obtained for streptomycin, novobiocin and neomycin, respectively. These catalytic surfaces can be renewed (by polishing), with a surface-to-surface reproducibility of 6.5% for the detection of 5 x 10(-5) M streptomycin. The analytical application of RuO2-modified carbon paste electrodes to the analysis of these antibiotics by flow injection was investigated, with a view to liquid chromatographic separation with electrochemical detection applications.

Catalysis↗

Determination of chloride at picogram levels by molecular fluorescence in a graphite furnace.

Chloride was determined at nanogram levels by adding excess of indium to the sample introduced into a graphite furnace and measuring the laser induced molecular fluorescence of indium chloride. The diatomic molecules of indium chloride were excited by a pulsed dye laser at 267 nm and fluorescence was measured at 359 nm. The effects of various parameters including amount of indium added, furnace thermal conditions and presence of concomitants were also studied. A linear calibration in the range of 0.025-1.25 ng and a detection limit of 17 pg of chloride were obtained under optimum conditions. The analytical usefulness of the method was checked by determining the chloride content in National Institute of Standards and Technology, Standard Reference Materials 1571a and 1571b Orchard Leaves.

Chlorides↗

Determination of lanthanum in food and water samples by Zeeman-effect atomic absorption spectrometry using graphite tube lined with tungsten foil.

A sensitive, selective method for the determination of lanthanum in food and water samples by atomic absorption spectrometry using a graphite tube lined with tungsten foil is described. The atomization of lanthanum from the tungsten surface gives better analytical sensitivity, a lower atomization temperature and negligible memory effects. The characteristic mass and detection limit of the method were 8.1 x 10(-9) and 7.85 x 10(-9) g, respectively. The precision (relative standard deviation in the range 5.9-9.9%), accuracy and interferences of the method were also investigated. The method can be used directly for the determination of trace amounts of lanthanum in food and water without pre-dissociation of the matrices. The results obtained by this method are in good agreement with those obtained from inductively coupled plasma atomic emission spectrometry.

Food Analysis↗

Renewable amperometric immunosensor based on paraffin-graphite-transferrin antiserum biocomposite for transferrin assay.

A renewable electrochemical immunosensor was developed for the determination of transferrin in human serum. It is based on a paraffin-graphite-transferrin antiserum biocomposite, which needs no additional curing. A competitive binding assay was used to determine transferrin in human serum with the aid of transferrin labeled with horseradish peroxidase. The assay conditions were optimized, including the loading of transferrin antiserum in the biocomposite, the amount of labeled transferrin in the incubation solution, incubation time and temperature. Serum samples were analyzed and the results demonstrate that the concentration range of determination with this system meets the demands of clinical analysis. The surface of the immunosensor can be regenerated by simply polishing to obtain a fresh immunocomposite ready to be used in a new competitive assay.

Biosensing Techniques↗

Multiresidue analysis of pesticides in vegetables and fruits using two-layered column with graphitized carbon and water absorbent polymer.

A high-throughput multiresidue analysis of pesticides in non-fatty vegetables and fruits was developed. The method consisted of a single extraction and a single clean-up procedure. Food samples were extracted with ethyl acetate and the mixture of extract and food dregs were poured directly into the clean-up column. The clean-up column consisted of two layers of water-absorbent polymer (upper) and graphitized carbon (lower), which were packed in a reservoir (75 ml ) of a cartridge column. The polymer removed water in the extract while the carbon performed clean-up. In a recovery test, 110 pesticides were spiked and average recoveries were more than 95% from spinach and orange. Most pesticides were recovered in the range 70-115% with RSD usually < 10% for five experiments. The residue analyses were performed by the extraction of 12 pesticides from 13 samples. The two methods resulted in similar residue levels except chlorothalonil in celery, for which the result was lower with the proposed method. The results confirmed that the proposed method could be applied to monitoring of pesticide residue in foods.

Food Contamination↗