Direct synthesis of graphene-chitosan composite and its application as an enzymeless methyl parathion sensor.

This paper proposed a direct electrodeposition approach to synthesis of graphene-chitosan (GR-CS) composite onto glassy carbon electrode (GCE) under controlled potential. This direct electrodeposition approach for the construction of GR-based hybrid was environmentally friendly, which would not involve the chemical reduction of graphene oxide (GO) and therefore result in no further contamination. The whole procedure was simply and cost only several minutes. Moreover, Combining the advantages of GR (large surface-to-volume ratio and high conductivity) and CS (good biocompatibility and adsorption), the GR-CS composite could be highly efficient to capture OPs and used as solid phase extraction (SPE). The GR-CS/GCE was used to detect organophosphate pesticides (OPs), using methyl parathion (MP) as a model analyte. The significantly redox response of MP on the GR-CS/GCE was proved. The linear range was wide from 4.0 ng mL(-1) to 400 ng mL(-1), and a low detection limit of 0.8 ng mL(-1) for MP was achieved. Moreover, the proposed sensor exhibited high reproducibility, long-time storage stability and satisfactory anti-interference ability. The proposed GR-CS/GCE opens new opportunity for green, fast, simple, and sensitive analysis of OP compounds.

Application of multi-walled carbon nanotubes modified carbon ionic liquid electrode for electrocatalytic oxidation of dopamine.

A simple, sensitive, and reliable method based on a multi-walled carbon nanotubes (MWNTs) modified carbon ionic liquid electrode (CILE) has been successfully developed for determination of dopamine (DA) in the presence of ascorbic acid (AA). The acid-treated MWNTs with carboxylic acid functional groups could promote the electron-transfer reaction of DA and inhibit the voltammetric response of AA. Due to the good performance of the ionic liquid, the electrochemical response of DA on the MWNTs/CILE was better than that of other MWNTs modified electrodes. Under the optimum conditions a linear calibration plot was obtained in the range 5.0×10(-8) to 2.0×10(-4) mol L(-1) and the detection limit was 1.0×10(-8) mol L(-1).

Bioremediation of heavy metals by growing hyperaccumulaor endophytic bacterium Bacillus sp. L14.

Heavy metal bioremediation by a multi-metal resistant endophytic bacteria L14 (EB L14) isolated from the cadmium hyperaccumulator Solanum nigrum L. was characterized for its potential application in metal treatment. 16S rDNA analysis revealed that this endophyte belonged to Bacillus sp. The hormesis of EB L14 were observed in presence of divalent heavy metals (Cu (II), Cd (II) and Pb (II)) at a relatively lower concentration (10mg/L). Such hormesis was the side effect of abnormal activities increases of ATPase which was planned to provide energy to help EB L14 reduce the toxicity of heavy metals by exporting the cations. Within 24h incubation, EB L14 could specifically uptake 75.78%, 80.48%, 21.25% of Cd (II), Pb (II) and Cu (II) under the initial concentration of 10mg/L. However, nearly no chromium uptake was observed. The mechanism study indicated that its remediation efficiencies may be greatly promoted through inhibiting the activities of ATPase. The excellent adaptation abilities and promising remediation efficiencies strongly indicated the superiority of this endophyte in heavy metal bioremediation at low concentrations, which could be useful for developing efficient metal removal system.

Amperometric glucose biosensor with remarkable acid stability based on glucose oxidase entrapped in colloidal gold-modified carbon ionic liquid electrode.

A colloidal gold-modified carbon ionic liquid electrode was constructed by mixing colloidal gold-modified graphite powder with a solid room temperature ionic liquid n-octyl-pyridinium hexafluorophosphate (OPPF(6)). Glucose oxidase (GOD) was entrapped in this composite matrix and maintained its bioactivity well and displayed excellent stability. The effect conditions of pH, applied potential and GOD loading were examined. Especially, the glucose oxidase entrapped in this carbon ionic liquid electrode fully retained its activity upon stressing in strongly acidic conditions (pH 2.0) for over one hour. The proposed biosensor responds to glucose linearly over concentration range of 5.0x10(-6) to 1.2x10(-3) and 2.6x10(-3) to 1.3x10(-2) M, and the detection limit is 3.5x10(-6) M. The response time of the biosensor is fast (within 10s), and the life time is over two months. The effects of electroactive interferents, such as ascorbic acid, uric acid, can be significantly reduced by a Nafion film casting on the surface of resulting biosensor.

Fabrication of DNA functionalized carbon nanotubes/Cu(2+) complex by one-step electrodeposition and its sensitive determination of nitrite.

In this paper, DNA functionalized single-wall carbon nanotubes/Cu(2+) (DNA-CNTs/Cu(2+)) complex was one-step electrodeposited onto the glassy carbon electrode (GCE), which fabricated a DNA-CNTs/Cu(2+)/GCE sensor to detect nitrite. Cyclic voltammogram of DNA-CNTs/Cu(2+)/GCE showed a pair of well-defined redox peaks for Cu(2+)/Cu(+). Compared with DNA-CNTs/GCE and DNA-Cu(2+)/GCE, the prepared DNA-CNTs/Cu(2+)/GCE exhibited more excellent electrochemical properties. Thus, the prepared DNA-CNTs/Cu(2+)/GCE was proposed as nitrite sensor. The effects of Cu(2+), CNTs and DNA concentration in the mixture together with electrodeposition time and determination conditions such as applied potential, pH value on the current response of DNA-CNTs/Cu(2+)/GCE toward nitrite were optimized to obtain the maximal sensitivity. In addition, electrochemical experiments revealed that the modified electrode showed high electrocatalytic activity to the reduction of nitrite ion (NO(2)(-)). The linear range for the detection of NO(2)(-) was 3x10(-8) to 2.6x10(-3)M, and the response was very fast (less than 3s). A low detection limit of 3x10(-8)M (S/N=3) for NO(2)(-) was achieved.

Direct electrochemistry and electrocatalytic properties of hemoglobin immobilized on a carbon ionic liquid electrode modified with mesoporous molecular sieve MCM-41.

The direct electron transfer and electrocatalysis of hemoglobin (Hb) entrapped in the MCM-41 modified carbon ionic liquid electrode (CILE) were investigated by using cyclic voltammetry in 0.10 M pH 7.0 phosphate buffer solution (PBS). Due to its uniform pore structure, high surface areas and good biocompatibility, the mesoporous silica sieve MCM-41 provided a suitable matrix for immobilization of biomolecule. The MCM-41 modified CILE showed significant promotion to the direct electron transfer of Hb, which exhibited a pair of well defined and quasi-reversible peaks for heme Fe(III)/Fe(II) with a formal potential of -0.284 V (vs. Ag/AgCl). Additionally, the Hb immobilized on the MCM-41 modified carbon ionic liquid electrode showed excellent electrocatalytic activity toward H(2)O(2). The electrocatalytic current values were linear with increasing concentration of H(2)O(2) in a wide range of 5-310 microM and the corresponding detection limit was calculated to be 5 x 10(-8)M (S/N=3). The surface coverage of Hb immobilized on the MCM-41 modified carbon ionic liquid electrode was about 2.54 x 10(-9) molcm(-2). The Michaelis-Menten constant K(m)(app) of 214 microM indicated that the Hb immobilized on the modified electrode showed high affinity to H(2)O(2). The proposed electrode had high stability and good reproducibility due to the protection effect of MCM-41 and ionic liquid, and it would have wide potential applications in direct electrochemistry, biosensors and biocatalysis.

Biosorption of cadmium by endophytic fungus (EF) Microsphaeropsis sp. LSE10 isolated from cadmium hyperaccumulator Solanum nigrum L.

A novel technology to obtain highly efficient biosorbent from the endophytes of a hyperaccumulator is reported. This technology is more convenient than the traditional method of obtaining biosorbents by experimentally screening many types of biomass by trial and error. Using this technology, endophytic fungus (EF) LSE10 was isolated from the cadmium hyperaccumulator Solanum nigrum L. It was identified as Microsphaeropsis sp. When cultured in vitro, the biomass yield of this EF was more than twice that of none-endophytic fungus (NEF) Rhizopus cohnii. Subsequently, it was used as a biosorbent for biosorption of cadmium from the aqueous solution. The results showed that the maximum biosorption capacity was 247.5mg/g (2.2 mmol/g) which was much higher than those of other adsorbents, including biosorbents and activated carbon. Carboxyl, amino, sulphonate and hydroxyl groups on EF LSE10 surface were responsible for the biosorption of cadmium.

A third-generation hydrogen peroxide biosensor based on horseradish peroxidase immobilized on DNA functionalized carbon nanotubes.

In this paper, DNA functionalized SWCNTs were used to immobilize horseradish peroxidase (HRP) on glassy carbon (GC) electrode. Cyclic voltammetry showed that the direct electrochemistry of HRP immobilized on DNA-SWCNTs hybrids was achieved. The DNA interlayer between the SWCNTs and HRP could be used to keep the activity of HRP. Compared with HRP-SWCNTs/GC and HRP-DNA/GC electrodes, the prepared HRP-DNA-SWCNTs/GC electrode exhibited more excellent electrochemical properties. Thus, the prepared HRP-DNA-SWCNTs/GC electrode was proposed as a third-generation H(2)O(2) biosensor. The effect of pH and applied potential on the performance of the biosensor was discussed in detail. Under the optimal conditions, a wide linear range of the propose biosensor for the detection of H(2)O(2) was observed from 6.0x10(-7) to 1.8 x10(-3)M. The detection limit was found to be 3.0 x10(-7)M at a signal-to-noise ratio of 3. Furthermore, the proposed biosensor displayed rapid response, high stability, very good reproducibility and high sensitivity for the detection of H(2)O(2). Determination H(2)O(2) concentration in disinfector sample by the proposed biosensor also showed satisfactory result.

Kinetic study and mathematical modeling of chromium(VI) reduction and microorganism growth under mixed culture.

The kinetics of chromium(VI) reduction by Pseudomonas aeruginosa (P. aeruginosa) and Escherichia coli (E. coli) was studied under both pure and mixed cultures. Initially, the study of kinetics was performed in pure culture. It was observed that the growth of the two bacteria was both inhibited in the presence of chromium(VI). The maximum specific growth rate (mu( m )) of P. aeruginosa decreased from 2.3942 h(-1) (without Cr(VI)) to 1.8551 h(-1) (with Cr(VI)). Under the mixed culture, the growth of E. coli was inhibited by P. aeruginosa. The maximum specific growth rate (mu( m )) of E. coli decreased form 0.871 h(-1) (in pure culture) to 0.153 h(-1) (in mixed culture). When the concentration of each bacterium was 4.5 x 10(8) cells ml(-1), the half-velocity reduction rate constant (K (C)) and the maximum specific reduction rate constant (v (max)) of chromium(VI) were 80.05 mg chromium(VI) l(-1) and 3.674 mg chromium(VI) cells(-1) h(-1), respectively. The results showed that the simulation appeared in good agreement with the experimental data, supporting the series of mathematical models represented the bacteria growth and chromium(VI) reduction in both pure and mixed cultures usefully.

Electrodeposition of chitosan-ionic liquid-glucose oxidase biocomposite onto nano-gold electrode for amperometric glucose sensing.

A sensitive glucose biosensor was fabricated by electrodepositing chitosan-ionic liquid-glucose oxidase biocomposite onto nano-gold electrode. First, nano-gold electrode was constructed by electrochemically depositing gold nanoparticles onto a flat gold electrode surface. Then the nano-gold electrode was immersed in the bath containing p-benzoquinone (BQ), chitosan (CS), glucose oxidase (GOD) and ionic liquid (IL) for electrodeposition of enzymatic electrode. The proton consumption during electroreduction of BQ increased the local solution pH near the electrode surface and led to the deposition of CS hydrogel on the electrode surface. Co-deposition of GOD and IL with the CS hydrogel was achieved. The proposed biosensor exhibited a fast amperometric response (<5 s) to glucose. Under the optimal conditions, the proposed biosensor exhibited a high current sensitivity (14.33 microA mM(-1) cm(-2)), which was 2.8 times of the biosensor prepared by electrodepositing CS-IL-GOD biocomposite on flat gold electrode. The detection limit for glucose was 1.5 microM, which was 20-fold lower compared to the biosensor prepared on flat gold electrode. The linear range for glucose detection was wide from 3.0 microM to 9.0 mM. Moreover, the proposed biosensor exhibited high reproducibility, long-time storage stability and satisfactory anti-interference ability. The applicability of the proposed biosensor to serum samples analysis was also evaluated.

Kinetic and equilibrium studies for the adsorption process of cadmium(II) and copper(II) onto Pseudomonas aeruginosa using square wave anodic stripping voltammetry method.

A novel method for the simultaneous determination of cadmium(II) and copper(II) during the adsorption process onto Pseudomonas aeruginosa was developed. The concentration of the free metal ions was successfully detected by square wave anodic stripping voltammetry (SWASV) on the mercaptoethane sulfonate (MES) modified gold electrode, while the P. aeruginosa was efficiently avoided approaching to the electrode surface by the MES monolayer. And the anodic stripping peaks of Cd(2+) and Cu(2+) appear at -0.13 and 0.34V respectively, at the concentration range of 5-50 microM, the peak currents of SWASV present linear relationships with the concentrations of cadmium and copper respectively. As the determination of Cd(2+) and Cu(2+) was in real time and without pretreatment, the kinetic characteristics of the adsorption process were studied and all the corresponding regression parameters were obtained by fitting the electrochemical experimental data to the pseudo-second-order kinetic model. Moreover, Langmuir and Freundlich models well described the biosorption isotherms. And there were some differences in the amount of metal ion adsorbed at equilibrium (q(e)) and other kinetics parameters when the two ions coexisted were compared with the unaccompanied condition, which were also discussed in this paper. The proposed electrode system provides excellent platform for the simultaneous determination of trace metals in complex biosorption process.

Layer-by-layer assembled carbon nanotube films with molecule recognition function and lower capacitive background current.

Multilayer films of multiwalled carbon nanotubes (MWCNTs) with molecule recognition function were assembled on glassy carbon (GC) electrode with lower capacitive background current by two steps: first, MWCNTs interacted with beta-cyclodextrin (beta-CD) with the aid of sonication to form beta-CD-MWCNTs nanocomposite, then the beta-CD-MWCNTs nanocomposite was assembled on GC electrode using layer-by-layer (LBL) method based on electrostatic interaction of positively charged biopolymer chitosan and negatively charged MWCNTs. The assembled beta-CD-MWCNTs multilayer films were characterized by scanning electron microscopy (SEM) and cyclic voltammetry. The SEM indicated that the MWCNTs multilayer films with beta-CD were somewhat more compact than that of the MWCNTs multilayer films without beta-CD. The cyclic voltammetric results indicated that the assembled MWCNTs with beta-CD on GC electrode exhibited lower capacitive background current than the assembled MWCNTs without beta-CD. The MWCNTs multilayer films with beta-CD were studied with respect to the electrocatalytic activity toward dopamine (DA). Compared with the MWCNTs multilayer films without beta-CD, the MWCNTs multilayer films with beta-CD possesses a much lower capacitive background current and higher electrocatalytic activity in phosphate buffer, which was ascribed to the relatively compact three-dimensional structure of the MWCNTs multilayer films with beta-CD and the excellent molecule recognition function of beta-CD.

[Separation and identification of 5 glycosidic flavor precursors in tobacco by ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry].

A qualitative method for the identification of 5 main glycosidic flavor precursors in tobacco was developed by using ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI MS/MS) and gas chromatography-mass spectrometry (GC-MS). The glycosidic flavor precursors in tobacco were extracted with methanol, cleaned up with an XAD-2 column. The aglycones were later released by enzyme-mediated hydrolysis under the condition of pH 5. The 5 volatile aglycone moieties were identified by GC-MS standard spectra library. The precursor ions of glycosides were determined by using electrospray ionization mass spectrometry in negative ion mode, then the 5 glycosidic flavor precursors were identified by using product ion scan (MS2) finally, using UPLC-ESI MS/MS, separation and identification of 5 glycosidic flavor precursors were accomplished on an RP-C,8 column in the multiple reaction monitoring (MRM) mode by using methanol and acetic acid-ammonium acetate aqueous solution as eluent. This work lays a foundation for the analysis of glycosidic flavor precursors without the standards by using liquid chromatography-mass spectrometry.

Detection of cytochrome c at biocompatible nanostructured Au-lipid bilayer-modified electrode.

The present work describes the dectection of cytochrome c (cyt c) at biocompatible aurum (Au) nanoparticle-structured supported bilayer lipid membrane (sBLM) modified with anionic sites. Au nanoparticles were directly deposited through sBLM modified with lauric acid (LA) to build a hybrid device of nanoscale electrode array via potential cycling in 10 mM HAuCl4 solution containing 0.1 M KCl. The properties of Au nanoparticle-doped sBLM composite were then characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). Results indicated that Au nanoparticles grew in voids of the sBLM with size around 20 - 30 nm. With SWV, after optimization, the results of the experiments indicate that the currents of cyt c were linear functions of its concentrations over the range from 1.0 x 10(-7) to 3.2 x 10(-6) M and the limit of detection (LOD, S/N = 3) was 5 x 10(-8) M. The influences of several common base pairs, amino acids and metal ions on determination of cyt c via this Au nanoparticle-doped sBLM composite were relatively low in experiments, suggesting the excellent biocompatibility of this detection method.

A monitoring technique of piezoelectric impedance analysis used in tea polyphenols (TP) antimutagenic characteristics.

A new method was proposed for studying antimutagenic characteristics of tea polyphenols (TP) using piezoelectric impedance analysis during the Salmonella typhimurium strain TA100 growth process in the presence of mutagen 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (abbreviated NNK). Compared with the general method of antimutagenic investigation, the proposed method provided real-time, multidimensional response information about the antimutagenic characteristics during the bacterial growth process. The results showed that TP was allowed to be antimutagenic at the lowest concentration of 0.25 microg/ml, and the inhibitory effect of TP presented a linear relationship with its dosage in the range of 0.25-2.5 microg/ml. And the relationship between the bacterial growth kinetic parameters and the TP dosage was also obtained. The present method provided a new approach for studying microbial antimutagenic characteristics.

Simultaneous voltammetry determination of dihydroxybenzene isomers by poly-bromophenol blue/carbon nanotubes composite modified electrode.

A novel modified electrode was constructed by electropolymerization of bromophenol blue at a multi-walled carbon nanotubes modified glassy carbon electrode. The electrode developed was used for the simultaneous determination of the isomers of dihydroxybenzene in environmental samples using a voltammetry method. There was a linear relationship over the range 10(-6)-10(-4 )mol L(-1) of hydroquinone, catechol and resorcinol; the detection limits was 3 x 10(-7) mol L(-1). The constructed electrode showed excellent reproducibility and stability. Actual water samples were analyzed and satisfactory result was obtained.

Direct electrochemistry and electrocatalysis of hemoglobin entrapped in semi-interpenetrating polymer network hydrogel based on polyacrylamide and chitosan.

Semi-interpenetrating polymer network (semi-IPN) hydrogel based on polyacrylamide (PAM) and chitosan was prepared to immobilize redox protein hemoglobin (Hb). The Hb-PAM-chitosan hydrogel film obtained has been investigated by scanning electron microscopy (SEM) and UV-VIS spectroscopy. UV-VIS spectroscopy showed that Hb kept its secondary structure similar to its native state in the Hb-PAM-chitosan hydrogel film. Cyclic voltammogram of Hb-PAM-chitosan film-modified glass carbon (GC) electrode showed a pair of well-defined and quasi-reversible redox peaks for Hb Fe(III)/Fe(II), indicating that direct electron transfer between Hb and GC electrode occurred. The electron-transfer rate constant was about 5.51 s(-1) in pH 7.0 buffers, and the formal potential (E degrees ') was -0.324 V (vs. SCE). The dependence of E degrees ' on solution pH indicated that one-proton transfer was coupled to each electron transfer in the direct electron-transfer reaction. Additionally, Hb in the semi-IPN hydrogel film retained its bioactivity and showed excellent electrocatalytic activity toward H(2)O(2). The electrocatalytic current values were linear with increasing concentration of H(2)O(2) in a wide range of 5-420 microM. The unique semi-IPN hydrogel would have wide potential applications in direct electrochemistry, biosensors and biocatalysis.

Determination of trace thiocyanate with nano-silver coated multi-walled carbon nanotubes modified glassy carbon electrode.

Novel nano-silver coated multi-walled carbon nanotube composites were prepared and used to fabricate a modified electrode. The application of the nano-silver coated multi-walled carbon nanotube composites modified electrode for determination of trace thiocyanate is demonstrated for the first time. The influence of substrate, pH and interference of coexisting substances was investigated for response properties of the electrode. There was a linear relationship at the range 2.5 x 10(-9) to 5 x 10(-8) mol L(-1) and 5 x 10(-8) to 1 x 10(-6) mol L(-1) of thiocyanate with the decrement of anodic DPV peak currents. The limit of detection was 1 x 10(-9) mol L(-1)(S/N=3). The constructed electrode showed excellent reproducibility and stability. Actual urine and saliva samples of smoker and non-smoker were analyzed and satisfactory results were obtained. This method provides a new way to construct any electrode for biological and environmental analysis.

Kinetics parameter estimation for the binding process of salicylic acid to human serum albumin (HSA) with capacitive sensing technique.

A novel method for real-time investigating the binding interaction between human serum albumin (HSA) and salicylic acid with capacitive sensing technique was successfully proposed. HSA was immobilized on the surface of a gold electrode modified with an insulating poly (o-phenylenediamine) (o-PD) film and colloid Au nanoparticles layers. The bioactivity of HSA was remained and major binding sites were available because of the excellent biocompatibility of gold nanoparticles. The capacitance and interfacial electron resistance of the sensor were altered, owing to the binding of HSA to salicylic acid. The time courses of the capacitance change were acquired with capacitive sensing technique during the binding process. Based on the capacitance response curves with time, the response model for the binding was derived in theory and the corresponding regression parameters were determined by fitting the real-time experimental data to the model. The binding and the dissociation rate constants (k(1) and k(-1)) were estimated to be 54.8 (mol l(-1))(-1) s(-1) and 2.9 x 10(-3) s(-1), respectively. And the binding equilibrium constant (K(a)) was calculated to be 1.89 x 10(4) (mol l(-1))(-1).

Immobilization of bovine serum albumin as a sensitive biosensor for the detection of trace lead ion in solution by piezoelectric quartz crystal impedance.

A biosensor based on bovine serum albumin (BSA) for the detection of lead (Pb(2+)) ion was developed and characterized. BSA was immobilized onto a colloidal Au-modified piezoelectric quartz crystal (PQC) as a biosensor for the detection of Pb(2+) ion by piezoelectric quartz crystal impedance (PQCI). Calibration curves for the quantification of Pb(2+) ion showed excellent linearity throughout the concentration range from 1.0 x 10(-7) to 3.0 x 10(-9)mol/L. The interaction between the Pb(2+) ions and the sensor chip is influenced significantly by the pH of the reaction buffer, and the optimal pH for the experiment was 5.4. Under the optimal conditions, the detection limit of 1.0 x 10(-9)mol/L for Pb(2+) was obtained. Kinetic parameters of the Pb(2+)-BSA interactions were also determined by using this chip. The sensor chip could be regenerated for use by dipping in the ethylenediaminetetraacetic acid (EDTA) solution for approximately 2h, and the chip was used to detect Pb(2+) ion for eight times without obvious signal attenuation.