Cysteine enhances clastogenic activity of dimethylarsinic acid

The effects of cysteine on dimethylarsinic acid (DMA)‐induced cytotoxicity and chromosomal aberration were studied using Chinese hamster V79 cells. The IC 50 of DMA, i.e. the concentration resulting in a 50% decrease in cell population of viable cells, was 130 µg ml^?1 (0.94 mM ), whereas that in the presence of 50 µg ml^?1 (0.28 mM ) cysteine was 20 µg ml^?1 (0.14 mM ). The mitotic index with co‐administration of 50 µg ml^?1 (0.36 mM ) DMA and 50 µg ml^?1 cysteine was 1.4 times that with 50 µg ml^?1 DMA alone. Whereas 82% of cells divided twice with 25 µg ml^?1 (0.18 mM ) DMA alone, most cells divided only once with co‐administration of 25 µg ml^?1 DMA and 50 µg ml^?1 cysteine. These results indicated that the increase in mitotic index by cysteine was due to enhancement of mitotic arrest by DMA. With co‐administration of 25 µg ml^?1 DMA and 50 µg ml^?1 cysteine, tetraploidy was 14.3% higher and fivefold by that with 25 µg ml^?1 DMA only. Cysteine at 50 µg ml^?1 enhanced induction of chromosomal aberrant cells by DMA. 100 µg ml^?1 (0.72 mM ) DMA induced 91% chromosomal aberrant cells in the presence of cysteine, and 12% in the absence of cysteine. Chromatid breaks and chromatid gaps were the most frequent types of aberration induced by co‐administration of DMA and cysteine or DMA alone. Co‐administration of DMA and cysteine produced many attenuated chromosomal figures. The attenuated chromosomal figures always had several chromatid gaps and chromatid breaks. Our findings may provide clues to arsenic carcinogenesis in humans. Copyright © 2002 John Wiley & Sons, Ltd.     

Cytotoxicological aspects of organic arsenic compounds contained in marine products using the mammalian cell culture technique

Arsenobetaine, arsenocholine, trimethylarsine oxide and tetramethylarsonium iodide, which are contained in marine fishery products, were examined for their potencies on cell growth inhibition, chromosomal aberration and sister chromatid exchange (SCE). Arseno- betaine, the major water-soluble organic arsenic compound in marine animals, exhibited very low cytotoxicity towards mammalian cells. This compound showed no cell growth inhibition at a concentration of 10 mg cm⁻³ and the cytotoxicity was lower than 1/14 000th of that of sodium arsenite and 1/1600th of that of sodium arsenate towards BALB/c 3T3 cells. The chromosomal aberrations caused by arsenobetaine at a concentration of 10 mg cm⁻³ consisted mainly of chromatid gaps and chromatid breaks, but in this concentration chromosomal breakage owing to its osmotic pressure is likely to be considerable. No SCE was observed at a concentration of 1 mg cm⁻³. Arsenocholine and trimethylarsine oxide also showed no cell growth inhibited at a concentration of 10 mg cm⁻³. However, tetramethylarsonium iodide inhibition the growth of BALB/c 3T3 at a concentration of 8 mg cm⁻³. These compounds exhibited a low ability to induce chromosomal aberrations at a concentration range of 2–10 mg cm⁻³ and no SCE was observed at a concentration of 1.0 mg cm⁻³. These results suggested that the major and minor organic arsenic compounds contained in marine fishery products are much less cytotoxic inorganic arsenic, methylarsonic acid and dimethylarsinic acid. © 1998 John Wiley & Sons, Ltd.     

磁性水铁矿的制备及其对二甲基亚胂酸的吸附性能

二甲基亚胂酸会对人体和环境造成严重危害。用水热法合成磁性水铁矿,对产物进行了X射线衍射分析、BET比表面积分析和磁滞回线分析,结果表明磁性水铁矿纯度较高,比表面积较大,具有较强的磁性。用磁性水铁矿作为吸附剂,考察了二甲基亚胂酸在磁性水铁矿上的吸附动力学及吸附等温线。二甲基亚胂酸在磁性水铁矿上的吸附符合准二级动力学模型,吸附速率为0.34g·mg^-1·h^-1;吸附等温线符合Freundlich模型。采用Zeta电位测定、FT-IR、SEM-EDS和XPS对吸附机理进行分析,表明磁性水铁矿通过配位络合作用和静电作用来吸附二甲基亚胂酸,在吸附过程中磁性水铁矿表面形了成Fe-O-As三元络合物。研究结果为含有二甲基亚胂酸污染物的水体处理和净化提供了新方法。     

Dimethylarsinic acid targets tubulin in mitotic cells to induce abnormal spindles

Dimethylarsinic acid (DMA) is the most effective inducer of cell‐cycle disruption among the arsenic compounds and their metabolites. The present study was conducted to gain further insight into cell‐cycle disruption induced by DMA. The inhibition of cell proliferation and the mitotic arrest induced by DMA were significant and dose‐dependent in Chinese hamster V79 cells and the two seemed to be closely related. At less than 140 µM the DMA did not inhibit the proliferation of cells, but it significantly induced mitotic arrest. An indirect immunofluorescence assay using anti‐α‐tubulin antibodies revealed that DMA induced the formation of abnormal spindles in the metaphase cells even at 350 µM with 5 h of treatment. At 1.4 mM DMA no metaphase cells could form a definite spindle structure. The spindle figures were similar to those induced by colchicine (125 nM ) or vinblastine (110 nM ), major antimitotic agents. In DMA‐treated interphase cells, the microtubule networks were indistinguishable from those of normal cells. With the tubulin‐assembly assay estimated by turbidity, DMA at less than 200 µM suppressed tubulin assembly in a dose‐dependent manner, whereas at more than 700 µM it enhanced tubulin polymerization remarkably with or without addition of excess guanosine‐5′‐triphosphate. According to the above findings, we discussed the possibility that DMA, a primary metabolite of inorganic arsenic in mammals, is related to arsenic carcinogenicity. Copyright © 2001 John Wiley & Sons, Ltd.     

The separation of dimethylarsinic acid,methylarsonous acid,methylarsonic acid,arsenate and dimethylarsinous acid on the Hamilton PRP‐X100 anion‐exchange column

In order to separate the potential arsenite metabolites methylarsonous acid and dimethylarsinous acid from arsenite, arsenate, methylarsonic acid and dimethylarsinic acid, the pH‐dependent retention behaviour of all six arsenic compounds was studied on a Hamilton PRP‐X100 anion‐exchange column with 30 mM phosphate buffers (pH 5, 6, 7, 8 and 9) containing 20% (v/v) methanol as mobile phase and employing an inductively coupled plasma atomic emission spectrometer (ICP–AES) as the arsenic‐specific detector. Baseline separation of dimethylarsinic acid, methylarsonous acid, methylarsonic acid, arsenate and dimethylarsinous acid was achieved with a 30 mmol dm⁻³ phosphate buffer (pH 5)–methanol mixture (80:20, v/v) in 25 min. Arsenite is not baseline‐separated from dimethylarsinic acid under these conditions. Copyright © 1999 John Wiley & Sons, Ltd.     

Induction of mitotic arrest and aneuploidy by organic arsenic compounds in human lymphocytes

Inorganic arsenic is methylated in the mammalian body to methylarsonic acid (MMA), dimethylarsinic acid (DMA) and trimethylarsine oxide (TMA). To achieve a more precise understanding of arsenic carcinogenicity, we examined the genotoxic effects of organic arsenic compounds on human lymphocytes by assessing induction of mitotic arrest, sister chromatid exchange (SCE) and aneuploidy. MMA, DMA and TMA arrested mitosis, DMA induced hyperdiploid cells, and DMA and TMA induced tetraploid cells. Of the three arsenic metabolites tested, DMA had the strongest effects on cell mitosis and aneuploidy induction. DMA arrested mitosis and induced c-mitosis significantly. These results suggest that DMA arrests mitosis and induces aneuploidy through spindle disruptions similar to those observed with known spindle poisons, such as colchicine or vinblastine. Since aneuploidy has been thought to be associated with tumor induction or neoplastic transformation, induction of aneuploidy by organic metabolites of arsenic may play a major role in arsenic carcinogenesis in humans. © 1997 John Wiley & Sons, Ltd.     

Analysis of Insulin by High Performance Liquid Chromatographic Method with Precolumn Derivatization with 4‐Chloro‐7‐Nitrobenzo‐2‐Oxa‐1,3‐Diazole

Abstract

A high performance liquid chromatographic method (HPLC) with precolumn derivatization and fluorescence detection for insulin was developed and applied for the quantification of insulin in spiked serum. To covalence couple with insulin, 4‐chloro‐7‐nitrobenzo‐2‐oxa‐1,3‐diazole (NBD‐Cl) was selected as fluorescent reagent. The optimal derivatization conditions were as follows: temperature 50; time 2 h, in the dark; 0.1 M phosphate buffer (pH 9.0). Analytical separation was carried out on a C18 column and the mobile phase including acetonitrile‐water containing 0.1% trifluoroacetic acid (TFA) (v/v∶ 30/70). The excitation/emission wavelengths were 470/540 nm. Under the conditions, the retention time and capacity factor of the adduct of insulin‐NBD were 10.03 min (flow rate 1 mL/min) and 3, respectively. The recovery of insulin in serum was 95.06% and the detection limit was 90 nM. In the investigated concentration ranges (0.46 µM～16.10 µM), R² was 0.9934, which indicated the potential for the application of NBD‐Cl derivatization to the analysis of insulin in the biological matrices, although with the shortcoming of long analytical time.     

Electrocatalytic Oxidation and Determination of Hydrazine at an AuCu Nanoparticles – Graphene – Ionic Liquid Composite Film Coated Glassy Carbon Electrode

Gold‐copper alloy nanoparticles (AuCu NPs) were electrodeposited on a graphene – ionic liquid composite film (EGN‐IL). The AuCu NPs showed high electrocatalysis to the oxidation of hydrazine with a catalytic reaction rate constant of about 5.0×10⁴ mol/Ls. In phosphate buffer solutions (pH 6.8) the oxidation current of hydrazine at 0.15 V (vs. SCE) at the resulting electrode (AuCu? EGN‐IL/GCE) was linear to its concentration in the range of 0.2–110 µM with a sensitivity of 56.7 µA/mM, and the detection limit was 0.1 µM (S/N=3). The electrode was successfully applied to the determination of waste water.     

Some critical aspects in the determination of binding constants by electrospray ionisation mass spectrometry at the example of arsenic bindings to sulphur‐containing biomolecules

The influences of reactant concentrations, solvent type, acid strength, pH conditions and ionic strength on the determination of apparent gas‐phase equilibrium constants K using electrospray ionisation mass spectrometry (ESI‐MS) were elucidated. As example serves the interaction of the tripeptide glutathione (GSH) with phenylarsine oxide (PAO). It was shown that rising initial concentrations of both reactants were not adequately compensated by increasing signal intensities of the reaction products in the mass spectra. The equilibrium constant for the formation of the phenylarsenic‐substituted peptide species decreased from 1.42 × 10⁵ ± 1.81 × 10⁴ l µmol^?1 to 1.54 × 10⁴ ± 1.5 × 10³ l µmol^?1 with rising initial GSH concentrations from 1 to 10 µM at fixed PAO molarity of 50 µM . K values resulting from a series with a fixed GSH molarity of 5 µM and a PAO molarity varied from 10 to 100 µM remained in a narrower range between 4.59 × 10⁴ ± 2.15 × 10⁴ l µmol^?1 and 1.07 × 10⁴ ± 4.0 × 10³ l µmol^?1. In contrast, consumption numbers calculated from the ion intensity ratios of reaction products to the unreacted peptide were not influenced by the initial reactant concentrations. In a water–acetonitrile–acetic acid mixture (48:50:2, v:v), the consumption of 5 µ M GSH increased from 8.3 ± 1.4% to 39.6 ± 1.6% with increased molar excess of PAO from 2 to 20, respectively. The GSH consumption was considerably enhanced in a changed solvent system consisting of 25% acetonitrile and 75% 10 mM ammonium formate, pH 5.0 (v:v) up to 80% of the original peptide amount at an only threefold molar arsenic excess. Copyright © 2010 John Wiley & Sons, Ltd.     

Amperometric Biosensors for Glucose Based on Layer‐by‐Layer Assembled Functionalized Carbon Nanotube and Poly (Neutral Red) Multilayer Film

Abstract

Multiwalled carbon nanotubes (MWNTs) were treated with a mixture of concentrated sulfuric and nitric acid to introduce carboxylic acid groups to the nanotubes. Conducting polymer film was prepared by electrochemical polymerization of neutral red (NR). By using a layer‐by‐layer method, homogeneous and stable MWNTs and poly (neutral red) (PNR) multilayer films were alternately assembled on glassy carbon (GC) electrodes. With the introduction of PNR, the MWNTs/PNR multilayer film system showed synergy between the MWNTs and PNR, with a significant improvement of redox activity due to the excellent electron‐transfer ability of carbon nanotubes (CNTs) and PNR. The electropolymerization is advantageous, providing both prolonged long‐term stability and improved catalytic activity of the resulting modified electrodes. The MWNTs/PNR multilayer film modified glassy carbon electrode allows low potential detection of hydrogen peroxide with high sensitivity and fast response time. As compared to MWNTs and PNR‐modified GC electrodes, the magnitude of the amperometric response of the MWNTs/PNR composite‐modified GC electrode is more than three‐fold greater than that of the MWNTs modified GC electrode, and nine‐fold greater than that of the PNR‐modified GC electrode. With the immobilization of glucose oxidase onto the electrode surface using glutaric dialdehyde, a biosensor that responds sensitively to glucose has been constructed. In pH 6.98 phosphate buffer, nearly interference‐free determination of glucose has been realized at ?0.2 V vs. SCE with a linear range from 50 µM to 10 mM and response time <10s. The detection limit was 10 µM glucose (S/N=3).     

Electrodeposition of PdAu Alloy Nanoparticles on Ionic Liquid Functionalized Graphene Film for the Voltammetric Determination of Oxalic Acid

An ionic liquid functionalized graphene film was prepared and PdAu nanoparticles (NPs) were electrodeposited on it. The PdAu NPs were characterized by various methods and they showed the features of alloys. In 0.2 M H₂SO₄ solution, oxalic acid (OA) exhibited a sensitive anodic peak at the resulting electrode at about 1.1 V (vs. SCE), and the peak current was linear to OA concentration in the range of 5–100 µM with a sensitivity of 45.5 µA/mM. The detection limit was 2.7 µM (S/N=3). The electrode was successfully applied to the determination of OA in real sample.     

Poly(pyridine‐3‐boronic acid)/Multiwalled Carbon Nanotubes Modified Glassy Carbon Electrodes for Simultaneous Determination of Ascorbic Acid, 3,4‐Dihydroxyphenylacetic Acid and Uric Acid

Poly(pyridine‐3‐boronic acid) (PPBA)/multiwalled carbon nanotubes (MWCNTs) composite modified glassy carbon electrode (GCE) was used for the simultaneous determination of ascorbic acid (AA), 3,4‐dihydroxyphenylacetic acid (DOPAC) and uric acid (UA). The anodic peaks for AA, DOPAC and UA at the PPBA/MWCNTs/GCE were well resolved in phosphate buffer solution (pH 7.4). The electrooxidation of AA, DOPAC and UA in the mixture solution was investigated. The peak currents increase with their concentrations increasing. The detection limits (S/N=3) of AA, DOPAC and UA are 5 µM, 3 µM and 0.6 µM, respectively.     

Isolation and evaluation of cytogenetic effect of Brahmi saponins on cultured human lymphocytes exposed in vitro

Major saponins of Brahmi (Bacopa monniera, Fam: Scrophulariaceae) – bacosides A and B – were isolated from the total methanol extract and characterised based on melting point, TLC, IR, ¹H NMR and ¹³C NMR. They were evaluated for their in vitro cytogenetic effects on human peripheral blood lymphocytes by chromosomal aberration (CA) assay and sister chromatid exchange (SCE) assay. The frequency of chromatid type aberrations and reciprocal interchanges between sister chromatids in the treated cells was scored in comparison to the untreated control. At 30 μg/mL dose, bacoside A showed a statistically significant increase in the frequency of both CA and SCE and bacoside B showed an increase only in SCE. Our report of the genotoxicity of the saponins is significant in view of the reports of anticancer activity of Brahmi extracts.     

Electrochemical Determination of Phenolic Acids at a Zn/Al Layered Double Hydroxide Film Modified Glassy Carbon Electrode

A stable sensor for the determination of gallic acid (GA) and caffeic acid (CA) was fabricated by electrodeposition of Zn‐Al‐NO₃ layered double hydroxide film on a glassy carbon electrode (LDHf/GCE). A sensitive electrochemical method was achieved for the determination of GA and CA in a phosphate buffer solution (pH 3). The differential pulse voltammetry response of the LDHf/GCE to GA has a linear concentration range from 4 µM to 600 µM with a correlation coefficient of 0.9985 and the calculated detection limit of 1.6 µM at a signal‐to‐noise ratio of 3. The differential pulse voltammetry response of the LDHf/GCE to CA has a linear concentration range from 7 µM to 180 µM with a correlation coefficient of 0.9969 and the calculated detection limit of 2.6 µM at a signal‐to‐noise ratio of 3. The constructed sensor was applied to the determination of GA in commercial green tea samples.     

Electrocatalytic Determination of Traces of Hydrazine by a Glassy Carbon Electrode Modified with Palladium‐Gold Nanoparticles

A glassy carbon electrode modified with palladium/gold nanoparticles was successfully prepared by an electrodeposition process. It efficiently oxidizes hydrazine at a low overpotential of ?0.26 V versus SCE. The Pd‐AuNPs with an average size of 50–80 nm are uniformly dispersed at the GCE. The Pd‐AuNPs/GCE was used for determination of hydrazine in phosphate buffer solution of pH 7.0. The amperometric current response of the electrode was increased linearly over a hydrazine concentration of 0.1–500 µM with a limit of detection of 0.07 µM .The prepared hydrazine sensor exhibited high sensitivity, good selectivity reproducibility and long term stability.     

Gold Nanoparticle Modified Screen Printed Electrodes for the Trace Sensing of Arsenic(III) in the Presence of Copper(II)

Gold ensembles for the trace level sensing of arsenic(III) in the presence of copper(II) are reported. The gold ensembles are fabricated using citrate capped gold nanoparticles which are chemically synthesised in an aqueous solution with an aliquot of this simply cast onto an economical and disposable screen printed electrode. After drying at room temperature, the gold ensembles are ready for use. The gold ensembles are explored towards the sensing of arsenic(III) in the presence of copper(II) using anodic stripping voltammetry where the corresponding stripping peaks are well resolved and using this protocol it is possible to readily detect 3 µg L^?1 (3 ppb) with a detection limit of 0.4 µg L^?1 (0.4 ppb). Proof‐of‐concept is also shown for the sensing of arsenic(III) in a canal water sample. Given the low cost of the sensor and ease of fabrication, the gold ensembles hold promise for the sensing of arsenic(III) in water samples where copper(II) may be present.     

Preparation,Characterization and Electrochemical Application of Graphene‐metallic Nanocomposites

Three different Graphene‐Metallic (Graphene‐Me) nanocomposites – Graphene‐Silver (Graphene‐Ag), Graphene‐Gold (Graphene‐Au) and Graphene‐Platinum (Graphene‐Pt) nanocomposites – were prepared and characterized. The electrochemical performances of these nanocomposites were tested by incorporating them with glassy carbon paste electrode (GCPE) and used them in biofuel cells (BFC) and as amperometric xanthine biosensor transducers. Present work contains the first application of Graphene‐Au and Graphene‐Ag nanocomposite in BFCs and also first application of these Graphene‐Me nanocomposites in xanthine biosensors. Considering BFC, power and current densities were calculated as 2.03 µW cm^?2 and 167.46 µA cm^?2 for the plain BFC, 3.39 µW cm^?2 and 182.53 µA cm^?2 for Graphene‐Ag, 4.43 µW cm^?2 and 230.15 µA cm^?2 for Grapehene‐Au and 6.23 µW cm^?2 and 295.23 µA cm^?2 for Graphene‐Pt nanocomposite included BFCs respectively. For the amperometric xanthine biosensor linear ranges were obtained in the concentration range between 5 µM and 50 µM with the RSD (n=3 for 30 µM xanthine) value of 4.28 % for plain xanthine biosensor, 3 µM and 50 µM with the RSD (n=3 for 30 µM xanthine) value of 9.37 % for Graphene‐Ag, 5 µM to 20 µM with the RSD (n=3 for 5 µM xanthine) value of 9.00 % and 30 µM to 70 µM with the RSD (n=3 for 30 µM xanthine) value of 8.80 % for Grapehene‐Au and 1 µM and 70 with the the RSD (n=3 for 30 µM xanthine) value of 2.59 % for Grapehene‐Pt based xanthine biosensors respectively.     

Electroanalysis of Norepinephrine at Bare Gold Electrode Pure and Modified with Gold Nanoparticles and S‐Functionalized Self‐Assembled Layers in Aqueous Solution. Part II

Gold electrodes modified with S‐containing compounds and gold were used for determination of norepinephrine (NEP) in aqueous solution. A linear relationship between norepinephrine concentration and current response was obtained in the range of 0.1 µM to 600 µM with the detection limit ≤0.090 µM for the electrodes modified at 2D template and in the range of 0.1 µM to 700 µM with the detection limit ≤0.075 µM for the electrodes modified at 3D template. The results have shown that modified electrodes could clearly resolve the oxidation peaks of norepinephrine, ascorbic (AA) and uric acid (UA) with peak‐to‐peak separation enabling determination of NEP, AA and UA in the presence of each other.     

Electrochemical Behavior and Electroanalytical Determination of Indole‐3‐Acetic Acid Phytohormone on a Boron‐Doped Diamond Electrode

In this work, a boron‐doped diamond (BDD) electrode was used for the electroanalytical determination of indole‐3‐acetic acid (IAA) phytohormone by square‐wave voltammetry. IAA yielded a well‐defined voltammetric response at +0.93 V (vs. Ag/AgCl) in Britton–Robinson buffer, pH 2.0. The process could be used to determine IAA in the concentration range of 5.0 to 50.0 µM (n=8, r=0.997), with a detection limit of 1.22 µM. The relative standard deviation of ten measurements was 2.09 % for 20.0 µM IAA. As an example, the practical applicability of BDD electrode was tested with the measurement of IAA in some plant seeds.     

Electropolymerization of Thin Film Conducting Polymer and Its Application for Simultaneous Determination of Ascorbic Acid,Dopamine and Uric Acid

In this paper electropolymerization of a thin film of para‐phenylenediamine (PPD) is studied at glassy carbon electrode (GCE) in sulfuric acid media by cyclic voltammetry. The results showed that this polymer was conducting and had a reproducible redox couple in the potential region from 0.0 to 0.4 V in phosphate buffer solution. This modified GCE (p‐PPD‐GCE) was applied for simultaneous determination of ascorbic acid (AA), dopamine (DA) and uric acid (UA) using differential pulse voltammetry (DPV). The p‐PPD‐GCE in 0.1 M phosphate buffer solution (pH 5.0) separated the DPV signals of AA, DA and UA with sufficient potential differences between AA–DA and DA–UA and also enhanced their oxidation peak currents. The oxidation currents were increased from 2.0 to 2000.0 µM for AA, 10.0 to 1250.0 µM for DA and 50.0 to 1600.0 µM for UA. The detection limits were evaluated as 0.4, 1.0 and 2.5 µM for AA, DA and UA, respectively (S/N=3).