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PubMedDate not supplied

AFM and STM study of beta-amyloid aggregation on graphite.

Atomic force microscopy (AFM) and scanning tunneling microscopy (STM) have been employed in situ and ex situ to directly study the aggregation of beta-amyloid(1-42) (Abeta42) peptide on hydrophobic graphite. From in situ AFM images, Abeta42 peptides were seen to aggregate into the sheets that preferred to three orientations with characteristic 3-fold symmetry (Proc. Natl. Acad. Sci. USA 96 (1999) 3688). The sheets were formed by parallel narrow lines with a height of 0.8-1.0nm and a width of 12-14nm. The narrow lines looked like beaded chains and have a right-handed axial periodicity. The high-resolution ex situ AFM and STM images showed that some fibrils of beta-amyloid had a characteristic domain texture, indicating they were formed through the association of protofibrils and monomers. The fibril containing lateral associated filaments that exhibited right-handed twist was clearly observed in the STM image. These results provide important clues to study the detailed structure of beta-amyloid aggregates and the mechanism of the Abeta fibrils formation on hydrophobic surface.

Alzheimer Disease↗

PubMed1996

Electrical injuries from graphite fishing rods.

High tension electrical injuries have a high morbidity and mortality. Carbon fibre and graphite used in the manufacture of fishing rods gives them superconductor qualities. The resistance of a fishing rod with a carbon content of more than 70 per cent is very low, ranging from 0.9 to 10.5 k omega. Nine cases are reported, which teach an important lesson not only in the management of those severe injuries, but also in their prevention.

Adult↗

PubMed1998

Liquid chromatography of guanidino compounds using a porous graphite carbon column and application to their analysis in serum.

The retention mechanism of guanidino compounds on a porous graphitic carbon seemed to be mainly hydrophobic interaction, according to the retention factors in buffer solutions and the results of an analysis by computational chemical calculation using molecular mechanics (MM2). The baseline separation of ten guanidino compounds was achieved by the addition of a hydrophobic counterion. The retention mechanism may be dynamic ion-exchange. The stable system was applied to the analysis of guanidino compounds in serum from nephritic patients. The effluent was monitored by a post-column labeling detection method using ninhydrin. The detection limit of guanidino compounds was a few picomoles; however, that of creatinine was one hundredth of those of the other compounds. The reproducibilities of the peak height and area of the ten guanidino compounds using gradient elution were quite high, and the standard deviations were within a few percent (n=5), except for creatinine. The recovery of the compounds from serum was more than 90% (n=5). The reproducibility of retention times was within 1% (n=5).

Buffers↗

PubMed2000

Rapid liquid chromatographic method for simultaneous determination of tetracyclines antibiotics and 6-epi-doxycycline in pharmaceutical products using porous graphitic carbon column.

A rapid and selective high performance liquid chromatographic (HPLC) method has been developed for the separation and determination of five commercially used tetracyclines. The chromatography was performed on a porous graphitic carbon (PGC) column, using 0.05 M potassium phosphate buffer (pH 2.0)--acetonitrile (40 + 60) as the mobile phase and ultraviolet detection at 268 nm. The method permits the simultaneous determination of oxytetracycline, metacycline, chlortetracycline and doxycycline as well as the separation of one of their common impurities (6-Epi-doxycycline) in bulk powder and pharmaceutical preparations with detection limits of 0.5-2 microg ml(-1) and recoveries of 98.9-100.5%. Correlation coefficients for calibration curves in the range of 5-50 microg ml(-1) were greater than 0.999 for all tetracyclines. The within- and between-day precision was determined for both retention times and peak area. It is suggested that the proposed HPLC-PGC method should be used for routine quality control and dosage form assay of tetracyclines in pharmaceutical preparations. The chromatographic behaviour of the five tetracyclines was examined under variable mobile phase compositions, the results revealed that elution order and selectivity were dependent on the buffer agent used. Comparison between retentions obtained with PGC and with silica-based stationary phase (ODS), showed similar variations of the capacity factors with the mobile phase composition, but with a different elution order.

Chromatography, High Pressure Liquid↗

PubMed2003

Simultaneous determination of naproxen and related compounds by HPLC using porous graphitic carbon column.

A simple, selective and sensitive high performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of naproxen and its main degradation products such as 1-(6-methoxy-2-naphthyl) ethanol (MNE), 2-methoxy-6-ethyl naphthalene (MEN) and 2-acetyl-6-methoxy naphthalene (AMN). The separation of these compounds was achieved on porous graphitic carbon (PGC) column using tetrahydrofuran-methanol as the mobile phase, and the effluent from the column was monitored at 272 nm. At a flow rate of 1 ml min(-1), the retention time of the last eluting compound was less than 10 min. Correlation coefficient for calibration curves in the ranges 2-25 microg ml(-1) for all compounds studied were greater than 0.999. The sensitivity of detection is 0.05 microg l(-1) for naproxen, MNE and MEN and 0.20 microg ml(-1) for AMN. The reproducibility of the peak area of these compounds using isocratic elution were quite high, and the standard deviations (S.D.) were below 2% (n=5). The reproducibility of retention times of these compounds was within 1% (n=5). The proposed liquid chromatographic method was successfully applied to the analysis of commercially available naproxen sodium (NS) dosage forms with recoveries of 98.8-102%. A comparative study shows that the selectivity of these compounds on PGC column was different to that obtained with octadecyl silica (ODS) columns.

Carbon↗

PubMed1997

Development and validation of a general non-digestive method for the determination of palladium in bulk pharmaceutical chemicals and their synthetic intermediates by graphite furnace atomic absorption spectroscopy.

A simple, selective, sensitive, accurate and relatively inexpensive method for the determination of palladium in bulk pharmaceutical chemicals (BPC) and their synthetic intermediates by graphite furnace atomic absorption spectroscopy has been developed and validated. Sample preparation by direct dissolution of sample in 70% nitric acid is simple and effective without adverse effects. The limit of detection and the limit of quantitation of the method were determined to be 0.7 ppm and 2 ppm respectively in BPC.

Acetates↗

PubMed1997

Separation of oligomers of nonylphenyl ethylene oxide [correction of nonlphenylethylene oxide] on a porous graphitized carbon column.

The retention of nonylphenyl ethylene oxide oligomer surfactants was determined on a porous graphitized carbon (PGC) column using water--methanol mixtures as eluents. Linear correlations were calculated between the logarithm of the capacity factor (k') and the methanol concentration in the eluent. To test the validity of the hypothesis that in the case of homologous series of solutes the intercept and slope values are intercorrelated linear correlation was calculated between the two chromatographic parameters. To elucidate the role of the length of the polar ethylene oxide chain in the retention linear correlations were calculated between the chromatographic parameters and the number of ethylene oxide groups per molecule. Nonylphenyl ethylene oxide oligomers were well separated on the PGC column. Significant linear relationships were found between the corresponding chromatographic parameters indicating that the solutes behave as a homologous series of compounds. The retention of surfactants increased linearly with increasing number of ethylene oxide groups per molecule indicating hydrophilic interactions between the solutes and the surface of PGC support.

Chromatography, High Pressure Liquid↗

PubMed1998

Separation of steroidal drugs on porous graphitized carbon column.

The retention time of 11 steroidal drugs was determined on a porous graphitized carbon (PGC) column using water tetrahydrofuran mixtures as eluents. Linear correlations were calculated between the logarithm of the capacity factor (k') and the tetrahydrofuran concentration in the eluent. To find the molecular parameters significantly influencing the retention the intercept and slope values of the above relationship were correlated with the physicochemical characteristics of steroidal drugs using principal component analysis (PCA). Both the slope and intercept values show high differences proving that steroidal drugs can be successfully separated on a PGC column. Five principal components explained > 90% of the total variance. Calculations indicated that both electrostatic interactions and sterical conditions can influence the retention of steroidal drugs on PGC column.

Chemistry Techniques, Analytical↗

PubMed2003

Graphite-epoxy composites as a new transducing material for electrochemical genosensing.

The use of a rigid carbon-polymer composite material as an electrochemical transducer in hybridisation genosensors is reported. Graphite-epoxy composites (GEC) have an uneven surface where DNA can be adsorbed using a simple dry-adsorption procedure. Single-stranded-DNA binds strongly to GEC in a way that prevents the strands from self-associating, while permitting hybridisation with complementary DNA. Hybridisation has been detected through biotin-streptavidin interaction using a streptavidin conjugated to horseradish peroxidase. Non-specific adsorption onto GEC is almost non-existent even when the surface has not been treated by blocking reagents. The analytical signal obtained was higher when compared with other electrochemical genosensors. Results can be achieved in 150 min, and the detection limit is in the order of fmol. Additionally, surface regeneration is possible using a simple polishing procedure, allowing for multiple use. The new genosensor based on GEC fulfils the requirements desired for these devices: ease of preparation as dry-adsorption of DNA is very simple and easily automated, robustness, sensitivity, low cost of production, ease of miniaturisation and simple use and fast response. Additionally, it can be used for field measurements and can be produced as a genosensor kit. Also, this material can be implemented for screen-printing procedures for the mass production of genosensors. The utility of the genosensor based on GEC is also illustrated with the detection of a sequence related to novel determinant of beta-lactamase resistance in Staphylococcus aureus.

Adsorption↗

PubMed2003

A nitric oxide biosensor based on the multi-assembly of hemoglobin/montmorillonite/polyvinyl alcohol at a pyrolytic graphite electrode.

Direct electron transfer of hemoglobin (Hb) can be achieved in a Hb/montmorillonite (MMT)/polyvinyl alcohol multi-assembly at a pyrolytic graphite (PG) electrode. Accordingly, a novel nitric oxide (NO) biosensor is proposed. The reduction of NO is observed at a potential of approximately -783 mV (vs. SCE) at pH 5.5. At optimum pH, this biosensor shows a wide linear range of 1.0x10(-6)-2.5x10(-4) mol/l with a detection limit of 5.0x10(-7) mol/l. The sensor-to-sensor reproducibility is good consideringmacr; a relative standard deviation of 3.5% in five independent determinations at 5.0x10(-5) mol/l NO. The modified electrode is conveniently constructed and durable in long-term operations.

Bentonite↗

PubMed2002

Comparison of graphite, aluminum, and TransHab shielding material characteristics in a high-energy neutron field.

Space radiation transport models clearly show that low atomic weight materials provide a better shielding protection for interplanetary human missions than high atomic weight materials. These model studies have concentrated on shielding properties against charged particles. A light-weight, inflatable habitat module called TransHab was built and shown to provide adequate protection against micrometeoroid impacts and good shielding properties against charged particle radiation in the International Space Station orbits. An experiment using a tissue equivalent proportional counter, to study the changes in dose and lineal energy spectra with graphite, aluminum, and a TransHab build-up as shielding, was carried out at the Los Alamos Nuclear Science Center neutron facility. It is a continuation of a previous study using regolith and doped polyethylene materials. This paper describes the results and their comparison with the previous study.

Aluminum↗

PubMed2002

Scanning probe microscopic imaging of guanine on a highly oriented pyrolytic graphite electrode.

Guanine adsorbed onto a highly oriented pyrolytic graphite electrode was studied by MAC-Mode Atomic Force Microscopy (AFM), and the electrochemical behaviour of the guanine layer was investigated with Electrochemical AFM. Guanine adsorbs spontaneously, without forming a well-packed structure, into nucleation spots, which are stable with time and cover the surface uniformly and almost completely. The process of guanine adsorption and nucleation can be controlled and the effect of altering the exposure time and varying the potential was investigated.

Adsorption↗

PubMed2003

Determination of isoprostanes in urine samples from Alzheimer patients using porous graphitic carbon liquid chromatography-tandem mass spectrometry.

F2-isoprostanes (F2-iPs) comprise four classes of isomers produced non-enzymatically by free radical attack on arachidonic acid, a component of the cell membrane. This paper describes a new method for the quantification of F2-isoprostanes in urine samples from thoroughly diagnosed Alzheimer's disease (AD) patients. The sample pretreatment consisted of liquid extraction of 900 microl urine with diethyl ether, its subsequent evaporation, and finally, reconstitution in 50 microl water. Of this, 20 microl was injected into a HPLC system with a 15 mm x 1 mm porous graphitic carbon column coupled to a triple quadrupole mass spectrometer running in negative electrospray ionization mode. The F2-isoprostanes were separated in 15 min using a linear solvent gradient comprising water, methanol, acetonitrile and ammonium hydroxide at a pH of 9.5. The average recovery obtained was approximately 75%. The limit of detection (3S/N) was calculated for iPF2alpha-III to be 0.7 pg injected on column, corresponding to 0.1 nM. The average level of iPF2alpha was 241 +/- 163 pg/mg creatinine in the urine samples from AD patients (average +/- standard deviation). The corresponding control values were 216 +/- 101 pg/mg creatinine, i.e. no statistically significant difference was noticed. No correlation pattern specific to Alzheimer's disease was revealed by principal component analysis of the isoprostane peaks obtained either. The results from this study support earlier findings that levels of peripheral isoprostanes are not increased in patients with Alzheimer's disease.

Aged↗

PubMed1991

Multiresidue method for pesticides in drinking water using a graphitized carbon black cartridge extraction and liquid chromatographic analysis.

A general liquid-solid extraction procedure for the isolation of pesticides from groundwater and drinking water for high-performance liquid chromatography (HPLC) is presented. This simple and rapid procedure involved passing a 2-L sample through a 250-mg graphitized carbon black (Carbopack B) cartridge at a flow rate of 150-160 mL/min. By taking advantage of the presence of positively charged active centers on the Carbopack B surface, a stepwise elution system allowed the complete separation of base-neutral pesticides from acidic ones. After partial solvent removal, the components in the two fractions were separated and quantified by gradient elution, reversed-phase HPLC with ultraviolet (UV) detection. The performance of the Carbopack cartridge was compared with that of a 500-mg C-18 bonded silica cartridge. With the Carbopack cartridge, the grand mean measurement accuracy of the 35 pesticides considered was 95%. With the C-18 cartridge, the grand mean measurement accuracy of the analytes was 76%. Compared to the C-18 cartridge, additional advantages of using a Carbopack cartridge are that the extraction procedure is about 7 times shorter, no pH adjustment of the environmental sample is necessary for trapping acidic compounds, and one cartridge instead of two suffices to extract base-neutral and acidic pesticides, making the Carbopack cartridge more adaptable than the C-18 one for field use. The detection limits by this method of all the pesticides considered were between 0.003 and 0.07 micrograms/L.

Carbon↗

PubMed2001

Graphite-teflon composite bienzyme electrodes for the determination of cholesterol in reversed micelles. Application to food samples.

A bienzyme amperometric composite biosensor for the determination of free and total cholesterol in food samples is reported. Cholesterol oxidase and horseradish peroxidase, together with potassium ferrocyanide as a mediator, are incorporated into a graphite-70% Teflon matrix. The compatibility of this biosensor design with predominantly nonaqueous media allows the use of reversed micelles as working medium. The reversed micelles are formed with ethyl acetate as continuous phase (in which cholesterol is soluble), a 4% final concentration of 0.05 mol L(-1) phosphate buffer solution, pH 7.4, as dispersed phase, and 0.1 mol L(-1) AOT as emulsifying agent. Studies on the repeatability of the amperometric response obtained at +0.10 V, with and without regeneration of the electrode surface by polishing, on the useful lifetime of one single biosensor and on the reproducibility in the fabrication of different pellets illustrate the robustness of the biosensor design. Determination of free and total cholesterol in food samples such as butter, lard, and egg yoke was carried out, and the obtained results were advantageously compared with those provided by using a commercial Boehringer test kit.

Biosensing Techniques↗

PubMed2001

HPLC determination of cyanuric acid in swimming pool waters using phenyl and confirmatory porous graphitic carbon columns.

The chlorinated salts of cyanuric acid have found an important role in recreational swimming pool waters across the United States. Upon application to pool water, they can (1) release disinfectant chlorine or (2) stabilize the free available chlorine by acting as chlorine reservoirs in the form of cyanuric acid, preventing the photolytic destruction of residual chlorine by sunlight. Recommended levels of the cyanuric acid stabilizer are in the 10-100 mg/L concentration range according to the National Swimming Pool Foundation (San Antonio, TX). Two isocratic HPLC methods with UV detection (213 nm) employing phenyl and porous graphitic carbon (PGC) columns and phosphate buffer eluents (pH 6.7 and pH 9.1, respectively) were developed to accurately measure cyanuric acid in swimming pools. The two methods allowed fast separation and detection of the stabilizer in 4 (phenyl) and 8 (PGC) min. Both methods offered practical sensitivities with method detection limits of 0.07 (phenyl) and 0.02 mg/L (PGC). Neither one of the two methods required the use of sample cleanup cartridges. They exhibit chromatograms with excellent baseline stability enabling low-level quantitation. Most important, the PGC column had a useful lifetime of five months and 500 sample analyses/column. Eleven pool water samples were fortified with 4.8-50.0 mg/L stabilizer, and the average recovery was 99.8%. Finally, statistical analysis on the relative precisions of the two methods indicated equivalence at the 0.05 critical level.

Chromatography, High Pressure Liquid↗

PubMed2002

Voltammetric determination of underivatized oligonucleotides on graphite electrodes based on their oxidation products.

A new electrochemical method to determine underivatized oligonucleotides is developed. The electro-oxidation of the adenine moieties of adsorbed oligonucleotides at elevated potentials on pyrolytic graphite electrodes (PGE) in neutral or alkaline media gives rise to electroactive products strongly adsorbed on the electrode surface. The extent of the redox processes of these products, with formal potential close to 0 V (vs Ag /AgCl) at pH 10, correlates well with the amount of parent oligonucleotide. Various electrochemical techniques have been compared and applied to the detection of specific DNA sequences and synthetic homopolynucleotides. Detection limits of 2 and 10 ng for (dA)20 and a 21-mer sequence of HIV-1, respectively, have been achieved using sample volumes of 10 microL. Moreover, the adsorbed oxidized oligonucleotide shows electrocatalytic activity toward the oxidation of NADH. The capability of the new method to detect DNA hybridization is discussed.

Chlamydia trachomatis↗

PubMed2003

Electrochemical genosensor based on colloidal gold nanoparticles for the detection of Factor V Leiden mutation using disposable pencil graphite electrodes.

Electrochemical genosensors for the detection of the Factor V Leiden mutation from polymerase chain reaction (PCR) amplicons using the oxidation signal of colloidal gold (Au) is described. A pencil graphite electrode (PGE) modified with target DNA, when hybridized with complementary probes conjugated to Au nanoparticles, responded with the appearance of a Au oxide wave at approximately +1.20 V. Specific probes were immobilized onto the Au nanoparticles in two different modes: (a) Inosine-substituted probes were covalently attached from their amino groups at the 5' end using N-(3-dimethylamino)propyl)-N'-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (NHS) as a coupling agent onto a carboxylate-terminated l-cysteine self-assembled monolayer (SAM) preformed on the Au nanoparticles, and (b) probes with a hexanethiol group at their 5' phosphate end formed a SAM on Au nanoparticles. The genosensor relies on the hybridization of the probes with their complementary targets, which are covalently immobilized at the PGE surface. Au-tagged 23-mer capture probes were challenged with the synthetic 23-mer target, 131-base single-stranded DNA or denatured 256-base polymerase chain reaction (PCR) amplicon. The appearance of the Au oxidation signal shortened the assay time and simplified the detection of the Factor V Leiden mutation from PCR amplified real samples. The discrimination between the homozygous and heterozygous mutations was also established by comparing the peak currents of the Au signals. Numerous factors affecting the hybridization and nonspecific binding events were optimized. The detection limit for the PCR amplicons was found to be as low as 0.78 fmol; thus, it is suitable for point-of-care applications.

DNA↗