Electrochemical detection of low concentration chloropropanol using silver nanowire array electrodes in aqueous media

In this paper, we report a facile method to fabricate silver nanowire array electrodes (SNAE) with ultra-high detection sensitivity to chloropropanol in the aqueous solution. Silver nanowire arrays were assembled in conventional anodic alumina membranes (AAM) by electrochemical deposition. Subsequently, silver nanowire arrays with an aspect ratio of 5 approximately 6 were deposited on the bottom of AAM. After a complete removal of the AAM , the grown arrays were used as working electrodes in a three-electrode cell. The electrochemical activity of SNAE was tested in the 0.1 mol/L NaClO4 aqueous solution using chloropropanol as analyte by a cyclic voltammetry method. The results show that SNAE display a distinct reduction peak at -1.011 V (vs. SCE) for chloropropanol and the linear dependencies of current on chloropropanol concentration were obtained within the concentration range 1.8 x 10(-7) approximately 2 x 10(-6) mol/L. The detection limit of chloropropanol was 10(-9) mol/L, which is significantly lower than that of their bulk counterparts. In short, SNAE show great potential in the determination of trace chloropropanol.     

Scanning electrochemical microscopy of quinoprotein glucose dehydrogenase

The activity of immobilized glucose dehydrogenase (GDH), a typical PQQ-dependent quinoprotein, was studied qualitatively and quantitatively by scanning electrochemical microscopy (SECM). PQQ-dependent GDH is of interest because of its high activity and independence of dissolved oxygen in catalyzing the transfer of electrons from glucose to an electron mediator. Biotinylated glucose dehydrogenase was bound to streptavidin-coated paramagnetic beads (surface concentration > or = 1.8 x 10(-11) mol cm(-2)) which were deposited as microscopic microspots on a hydrophobic surface. The catalytic activity of immobilized GDH was mapped in SECM feedback mode and generation-collection mode using ferrocenemethanol, ferrocenecarboxylic acid, p-aminophenol, and ferricyanide as electron mediators, respectively. The apparent steady-state kinetics of catalysis were measured under conditions of high d-glucose concentration using the theory developed for the SECM feedback and generation collection (GC) modes. In feedback mode, curves of the kinetically controlled substrate current against normalized distance were plotted, and it was found that GDH catalysis follows pseudo-first-order kinetics. In GC mode detection, the catalysis follows zero-order kinetics in the presence of high concentration of both substrates for GDH. The turnover rate obtained for immobilized GDH is lower than that of native GDH but much higher than that generally observed for glucose oxidase.     

Analysis of phenothiazine and its derivatives using LC/electrochemistry/MS and LC/electrochemistry/fluorescence

The on-line electrochemical conversion of phenothiazine and its derivatives after liquid chromatographic separation has been studied by mass spectrometry and fluorescence spectroscopy. In an electrochemical cell consisting of porous glassy carbon, the phenothiazines are readily converted to oxidized products, which can be detected by on-line fluorescence spectroscopy and mass spectrometry. The method allows rapid investigations on the electrochemical oxidation pathways, as demonstrated for phenothiazine itself. The phenothiazine derivatives are transferred into their strongly fluorescent sulfoxides. Based on this reaction, an LC/electrochemistry/fluorescence method was developed that allows for limits of detection between 5 x 10(-9) and 4 x 10(-8) mol/L and limits of quantification between 2 x 10(-8) and 1 x 10(-7) mol/L for the individual phenothiazines. The linear ranges comprised three decades starting at the limit of quantification.     

Oxygen consumption of single bovine embryos probed by scanning electrochemical microscopy

Oxygen consumption of individual bovine embryos was noninvasively quantified by scanning electrochemical microscopy (SECM). A probe microelectrode was used to scan near a single embryo surface in a culture medium to monitor the oxygen reduction current at 37 degrees C, under a water-saturated atmosphere of 5% CO2 and 95% air. The oxygen concentration profiles near the embryos were in good agreement with the theoretical spherical diffusion. When an embryo reached the stage of a morula with a 74-microm radius on day 6 after in vitro fertilization, the oxygen concentration difference (deltaC) between the bulk solution and the morula surface was 6.90 +/- 1.35 microM. The oxygen consumption rate (F) of the single morula was estimated to be (1.40 +/- 0.27) x 10(-14) mol s(-1). After the SECM measurement, the embryo was continuously cultured for another 2 days and grew to the stage of a blastocyst with a 100-microm radius. For the blastocyst, the deltaC values for the inner cell mass side and the trophoblast side were 16.40 +/- 1.83 and 9.14 +/- 1.68 microM, respectively. The oxygen consumption rate of the blastocyst was found to be in the range of (2.50 +/- 0.46) x 10(-14) mol s(-1) < F < (4.49 +/- 0.50) x 10(-14) mol s(-1). We have carried out SECM measurements for 19 embryos, and the results were compared in detail with these from an optical microscopic observation. The deltaC values for the morulae on day 6 after in vitro fertilization were strongly related to the morphological embryo quality. The morulae showing a larger deltaC value developed into blastocysts of a larger size, and the deltaC value after the subsequent 2 days of cultivation was found to be increased.     

Scanning electrochemical microscopy. 40. Voltammetric ion-selective micropipet electrodes for probing ion transfer at bilayer lipid membranes

Voltammetric ion-selective micropipet electrodes for use in scanning electrochemical microscopy (SECM) for detection of potassium ion were fabricated. These used pulled borosilicate capillaries with tip orifice radii of 0.7-20 microm with silanized inner walls filled with a solution of 10 mM valinomycin and 10 mM ETH 500 in dichloroethane. The electrodes were characterized by determining the steady-state tip current for K+ concentrations of 0.05-0.3 mM. The tips were used in the SECM feedback and generation-collection modes to study K+ transfer through gramicidin channels in a horizontal bilayer lipid membrane (glycerol monooleate).     

Scanning electrochemical microscopy. 44. Imaging of horseradish peroxidase immobilized on insulating substrates

Scanning electrochemical microscopy (SECM) was used to study horseradish peroxidase (HRP) immobilized with copolymer on insulating substrates (glass slide or polycarbonate membrane filter). Two methods were used to immobilize HRP: In the first, HRP was coimmobilized by cross-linking on a glass slide with a copolymer swelled in water to form a hydrogel; in the second, the same copolymer and avidin were coimmobilized on the glass slide and biotin-labeled HRP was conjugated to the avidin of the film. SECM was then used to detect the presence of the bound enzyme by observing the feedback current in a solution of benzoquinone and hydrogen peroxide, when hydroquinone was generated at the tip. A detection limit less than 7 x 10(5) HRP molecules within a approximately 7-microm-diameter area was demonstrated.     

Analytical technique for label-free multi-protein detection based on Western blot and surface-enhanced Raman scattering

We have developed a new analytical procedure for label-free protein detection designated "Western SERS", consisting of protein electrophoresis, Western blot, colloidal silver staining, and surface-enhanced Raman scattering (SERS) detection. A novel method of silver staining for Western blot that uses a silver colloid, an excellent SERS-active substrate, is first proposed in the present study. During the process of silver staining, interactions between proteins and silver nanoparticles result in the emergence of SERS of proteins. In the present study, we use myoglobin (Mb) and bovine serum albumin (BSA) as model proteins. From different protein bands on a nitrocellulose (NC) membrane, we have observed surface-enhanced resonance Raman scattering (SERRS) spectra of Mb and SERS spectra of BSA. The proposed technique offers dual advantages of simplicity and high sensitivity. On one hand, after the colloidal silver staining, we can detect label-free multi-proteins directly on a NC membrane without digestion, extraction, and other pretreatments. On the other hand, the detection limit of the Western SERS is almost consistent with the detection limit of colloidal silver staining, and the SERRS detection limit of Mb is found to be 4 ng/band. This analytical method, which combines the technique of protein separation with SERS, may be a powerful protocol for label-free protein detection in proteomic research.     

Local feedback mode of scanning electrochemical microscopy for electrochemical characterization of one-dimensional nanostructure: theory and experiment with nanoband electrode as model substrate

Local feedback mode is introduced as a novel operation mode of scanning electrochemical microscopy (SECM) for electrochemical characterization of a single one-dimensional (1D) nanostructure, for example, a wire, rod, band, and tube with 1-100-nm width and micrometer to centimeter length. To demonstrate the principle, SECM feedback effects under diffusion limitation were studied theoretically and experimentally with a disk probe brought near a semi-infinitely long band electrode as a geometrical model for a conductive 1D nanostructure. As the band becomes narrower than the disk diameter, the feedback mechanism for tip current enhancement is predicted to change from standard positive feedback mode, to positive local feedback mode, and then to negative local feedback mode. The negative local feedback effect is the only feedback effect that allows observation of a 1D nanostructure without serious limitations due to small lateral dimension, available tip size, or finite electron-transfer rate. In line-scan and approach-curve experiments, an unbiased Pt band electrode with 100-nm width and 2.6-cm length was detectable in negative local feedback mode, even using a 25-microm-diameter disk Pt electrode. Using a 2-microm-diameter probe, both well-defined and defected sites were observed in SECM imaging on the basis of local electrochemical activity of the nanoband electrode. Noncontact and spatially resolved measurement is an advantage of this novel SECM approach over standard electrochemical approaches using electrodes based on 1D nanostructure.     

Detection of microspotted carcinoembryonic antigen on a glass substrate by scanning electrochemical microscopy

Microspots of carbinoembryonic antigen (CEA) on glass substrates were characterized by scanning electrochemical microscopy (SECM). CEA was immobilized via a sandiwch method using horseradish peroxidase (HRP)-labeled anti-CEA. The reduction current of the oxidized form of ferrocenylmethanol generated by the HRP reaction was monitored to view SECM images. This method detects as low as ～10(4) CEA molecules in a single 20-μm-radius spot.     

Distinction of the two phases of CuTCNQ by scanning electrochemical microscopy

The two known phases of CuTCNQ and TCNQ (TCNQ = 7,7',8,8'-tetracyanoquinodimethane) have been probed by scanning electrochemical microscopy (SECM) in the feedback mode. The first use of this technique for distinguishing differences in the electronic properties of semiconductor phases exploits the large differences in conductivity that exist between CuTCNQ and the parent TCNQ material and also between the CuTCNQ phases I and II. However, the packing density of the individual CuTCNQ crystals in a film structure also is shown to influence the SECM feedback response. Finally, it is shown that films of pure phase II material or mixtures of the phases can be mapped using feedback mode SECM. The SECM method provides valuable insights for elucidating properties of semiconducting solids that are mounted on insulating substrates.     

Carbon nanofiber electrodes and controlled nanogaps for scanning electrochemical microscopy experiments

The electrochemical behavior of electrodes made by sealing carbon nanofibers in glass or with electrophoretic paint has been studied by scanning electrochemical microscopy (SECM). Because of their small electroactive surface area, conical geometry with a low aspect ratio and high overpotential for proton and oxygen reduction, carbon nanofiber (CNF) electrodes are promising candidates for producing electrode nanogaps, imaging with high spatial resolution and for the electrodeposition of single metal nanoparticles (e.g., Pt, Pd) for studies as electrocatalysts. By using the feedback mode of the SECM, a CNF tip can produce a gap that is smaller than 20 nm from a platinum disk. Similarly, the SECM used in a tip-collection substrate-generation mode, which subsequently shows a feedback interaction at short distances, makes it possible to detect a single CNF by another CNF and then to form a nanometer gap between the two electrodes. This approach was used to image vertically aligned CNF arrays. This method is useful in the detection in a homogeneous solution of short-lifetime intermediates, which can be electrochemically generated at one electrode and collected at the second at distances that are equivalent to a nanosecond time scale.     

Alternating current impedance imaging of membrane pores using scanning electrochemical microscopy

Alternating current impedance imaging of a 6-microm thick membrane containing conical-shaped pores (60-nm and 2.5-microm diameter openings) using scanning electrochemical microscopy (SECM) is described. Impedance images of the pore openings were obtained by rastering a glass-sealed conically shaped Pt tip (approximately 1-microm radius) above the membrane surface, while measuring the total impedance between the tip and a large area Pt electrode located on the opposite side of the membrane. Individual pore openings in the high pore density membrane (approximately 8 x 10(4) pores/cm2) are observed in the SECM impedance image. The image contrast is due to the decrease in tip and membrane resistance, in the vicinity of the pore opening. An equivalent circuit for the SECM cell and membrane is proposed and evaluated against the measured SECM imaging impedance. Criteria for employing SECM in impedance mode to image membranes are discussed.     

Scanning electrochemical microscopy monitoring in microcantilever platforms

The deflection of cantilever systems may be performed by an indirect electrochemical method that consists of measuring the local cantilever activity and deflection in a feedback generation-collection configuration of the SECM. This is illustrated during the electrochemically assisted adsorption of Br onto a gold-coated cantilever, either in its pristine state or previously coated with a thin organic barrier. It is further extended to the adsorption of an antibody in a heterogeneous immunoassay at an allergen-coated microcantilever platform. In both reactions, the cantilever deflection is qualitatively detected from the SECM tip current measurement and a quantitative estimate is obtained through modeling. This electroanalytical strategy provides an alternative approach to standard optical detection. It can overcome some limitations of the optical method by allowing electrochemical characterization of nonconductive cantilevers and appropriate use for closed systems.     

Carbon fiber nanoelectrodes modified by single-walled carbon nanotubes

Microelectrode voltammetry has been considered to be a powerful technique for single biological cell analysis and brain research. In this paper, we have developed a simple method to get highly sensitive carbon fiber nanoelectrodes (CFNE) modified by single-walled carbon nanotubes (SWNTs) on the basis of our previous work. The electrochemical behavior of SWNTs/CFNE was characterized by potassium ferricyanide, dopamine (DA), epinephrine (E), and norepinephrine (NE) using cyclic voltammetry (CV). Compared with CFNE, SWNTs/CFNE has a much larger available internal surface area per external geometric area, which is supported by SEM images. The modified electrodes show very high sensitivity and favorable electrochemical behavior toward these neurotransmitters. The peak current increases linearly with the concentration of DA, E, and NE in the range of 1.0 x 10(-)(7)-1.0 x 10(-)(4), 3.0 x 10(-)(7)-1.0 x 10(-)(4), and 5.0 x 10(-)(7)-1.0 x 10(-)(4) M, respectively. The CV detection limit (S/N = 3) of DA, E, and NE is 7.7 x 10(-)(9), 3.8 x 10(-)(8), and 4.2 x 10(-)(8) M, respectively. The modified electrode exhibited almost the same electrochemical behavior after 15 days, indicating that SWNTs/CFNE is pretty stable and has good reproducibility.     

Fabrication of nano-porous hydroxyapatite modified electrode and its application for determination of p-chlorophenol

Nano-porous hydroxyapatite (HAp) modified electrode was fabricated by simply electrodepositing HAp onto the glassy carbon electrode (GCE) from the electrolytes solution containing Ca(NO3)2 4H2O and NH4H2PO4, the resulting electrode (nano-HAp/GCE) was characterized with scanning electron microscopy (SEM). The electrochemical behavior of p-chlorophenol (p-CP) at nano-HAp/GCE was studied by cyclic voltammetry. The electrode displayed selective and enhanced electroanalytical response towards p-CP, obviously because p-CP is accumulated at the electrode. For the greater sensitivity, a semi-derivative technique was adopted to obtain the current signal. The results indicated that the nano-HAp/GCE exhibits substantial enhancement in electrochemical sensitivity for p-CP due to its large surface area and particular adsorbability. After accumulation of 4 min for p-CP on nano-HAp/GCE, the peak height was linearly related to the concentration of p-CP in the range of 1.0 x 10(-8) to 1.0 x 10(-7) mol L(-1). The detection limit was 4.0 x 10(-9) mol L-(1) at 3sigma level. Based on this, the modified electrode was successfully applied in water samples with low cost and high sensitivity.     

石墨烯/银纳米颗粒复合材料的制备及其对双氧水的电化学检测

以氧化石墨烯(GO)和硝酸银为原材料,聚乙烯吡咯烷酮(PVP)为还原剂和稳定剂,通过水热法制备出还原氧化石墨烯/银纳米颗粒(rGO/AgNPs)复合材料。采用透射电子显微镜(TEM)、X射线衍射(XRD)及紫外-可见分光光度计(UV-Vis)对rGO/AgNPs复合材料的形貌、组成和结构进行表征。同时,将rGO/AgNPs复合材料修饰到玻碳电极表面制备出过氧化氢(H_2O_2)电化学传感器,通过循环伏安法(CV)和计时安培响应法(i-t)对传感器进行电化学性能测试。实验结果表明:制备的rGO/AgNPs传感器具有较好的电化学性能,其对H_2O_2检测的灵敏度为340.6μA·(mmol/L)~(-1)·cm~(-2),响应时间为3s,最低检测极限为7.5μmol/L(S/N=3),线性检测范围为20~4950μmol/L(线性相关系数为R=0.9973)。     

Combined scanning electrochemical/optical microscopy with shear force and current feedback

A technique that combines scanning electrochemical microscopy (SECM) and scanning optical microscopy (OM) was developed. Simultaneous scanning electrochemical/optical microscopy (SECM/OM) was performed by a special probe tip, which consists of an optical fiber core for light passage, surrounded by a gold ring electrode, and an outermost electrophoretic insulating sheath, with the tip attached to a tuning fork. To regulate the tip-substrate distance, either the shear force or the SECM tip current was employed as the feedback signal. The application of a quartz crystal tuning fork (32.768 kHz) for sensing shear force allowed simultaneous topographic, along with SECM and optical imaging in a constant-force mode. The capability of this technique was confirmed by obtaining simultaneously, for the first time, topographic, electrochemical, and optical images of an interdigitated array electrode. Current feedback from SECM also provided simultaneous electrochemical and optical images of relatively soft samples, such as a polycarbonate membrane filter and living diatoms in a constant-current mode. This mode should be useful in mapping the biochemical activity of a living cell.     

Fabrication and characterization of probes for combined scanning electrochemical/optical microscopy experiments

A technique that combines scanning electrochemical microscopy (SECM) and optical microscopy (OM) was implemented with a new probe tip. The tip for scanning electrochemicaVoptical microscopy (SECM/OM) was constructed by insulating a typical gold-coated near-field scanning optical microscopy tip using electrophoretic anodic paint. Once fabricated, the tip was characterized by steady-state cyclic voltammetry, as well as optical and electrochemical approach experiments. This tip generated a stable steady-state current and well-defined SECM approach curves for both conductive and insulating substrates. Durable tips whose geometry was a ring with < 1 microm as outer ring radius could be consistently fabricated. Simultaneous electrochemical and optical images of an interdigitated array electrode were obtained with a resolution on the micrometer scale, demonstrating good performance of the tip as both an optical and an electrochemical probe for imaging microstructures. The SECM feedback current measurements were successfully employed to determine tip-substrate distances for imaging.     

Photothermal deflection determination of iron(II) with ferrozine with sorption preconcentration on Silufol plates

Photothermal deflection spectroscopy was applied to the selective detection of iron(II) chelate with ferrozine by its sorption preconcentration on Silufol plates. The linearity range was 1 x 10(-11) - 6 x 10(-8) mol cm(-2) of chelate at the plate surface, which corresponded to 1 x 10(-9) -4 x 10(-6) M of chelate in solution. The limits of detection and quantification are 8 x 10(-12) and 2.5 x 10(-11) mol cm(-2) at the plate from 15 microL of test solution (0.5 nM and 1.5 nM in solution, respectively), and the absolute detection limit is 8 fmol in the whole spot applied to a plate. Characteristics and features of photothermal deflection detection are discussed.