Nasal Immunization Using Chitosan Nanoparticles with Glycoprotein B of Murine Cytomegalovirus.

The use of nanoparticles as a delivery system for a specific antigen could solve many limitations of mucosal vaccine applications, such as low immunogenicity, or antigen protection and stabilization. In this study, we tested the ability of nasally administered chitosan nanoparticles loaded with glycoprotein B of murine cytomegalovirus to induce an immune response in an animal model. The choice of chitosan nanoparticle type was made by in vitro evaluation of sorption efficiency and antigen release. Three types of chitosan nanoparticles were prepared: crosslinked with tripolyphosphate, coated with hyaluronic acid, and in complex with polycaprolactone. The hydrodynamic size of the nanoparticles by dynamic light scattering, zeta potential, Fourier transform infrared spectroscopy, scanning electron microscopy, stability, loading efficiency, and release kinetics with ovalbumin were evaluated. Balb/c mice were immunized intranasally using the three-dose protocol with nanoparticles, gB, and adjuvants Poly(I:C) and CpG ODN. Subsequently, the humoral and cell-mediated antigen-specific immune response was determined. On the basis of the properties of the tested nanoparticles, the cross-linked nanoparticles were considered optimal for further investigation. The results show that nanoparticles with Poly(I:C) and with gB alone raised IgG antibody levels above the negative control. In the case of mucosal IgA, only gB alone weakly induced the production of IgA antibodies compared to saline-immunized mice. The number of activated cells increased slightly in mice immunized with nanoparticles and gB compared to those immunized with gB alone or to negative control. The results demonstrated that chitosan nanoparticles could have potential in the development of mucosal vaccines.

Effect of different modification by gold nanoparticles on the electrochemical performance of screen-printed sensors with boron-doped diamond electrode.

Screen-printed sensors with chemically deposited boron-doped diamond electrodes (BDDE) were modified with different types of gold nanoparticles (AuNPs) according to a new original procedure. Physically and electrochemically deposited AuNPs had various sizes and also nanoporous character. They also differ in shape and density of surface coverage. The developed sensors were characterized using scanning electron microscopy and Raman spectroscopy. Their electrochemical properties were studied using cyclic voltammetry and electrochemical impedance spectrometry of selected outer sphere ([Ru(NH3)6]Cl3) and inner sphere (K3[Fe(CN)6], dopamine) redox markers. The application possibilities of such novel screen-printed sensors with BDDE modified by AuNPs were verified in the analysis of the neurotransmitter dopamine. The best analytical performance was achieved using printed sensors modified with the smallest AuNPs. The achieved limit of detection values in nanomolar concentrations (2.5 nmol L-1) are much lower than those of unmodified electrodes, which confirms the significant catalytic effects of gold nanoparticles on the surface of the working electrode. Sensors with the best electrochemical properties were successfully applied in the analysis of a model solution and spiked urine samples.

New electroanalytical method for the determination of trans-anethole in spices and sweets.

A completely new method for the determination of trans-anethole (TAN) based on the anodic oxidation of this flavouring substance in pure acetonitrile using differential-pulse voltammetry has been developed. A nonaqueous carbon paste electrode bulk-modified with solid sodium dodecyl sulphate of 40 % (w/w) content was chosen as optimum. To propose TAN electrode reaction mechanism, its electrochemical behaviour was investigated at glassy carbon electrode in nonaqueous media. At optimum working conditions, the current response could be calibrated within a linear range 2-200 µmolL-1 TAN, a coefficient of determination of 0.9971, a sensitivity of 0.1122µALµmol-1, and a detection limit of 0.7 µmolL-1. A satisfactory precision (relative standard deviation of 4 %) has been achieved. The validation performed by analysis of spices and sweets provided comparable results with reference reverse-phase HPLC with spectrophotometric detection, thus confirming the practical use of the developed voltammetric method in food analysis.

Screen-Printed Carbon Electrodes with Macroporous Copper Film for Enhanced Amperometric Sensing of Saccharides.

A porous layer of copper was formed on the surface of screen-printed carbon electrodes via the colloidal crystal templating technique. An aqueous suspension of monodisperse polystyrene spheres of 500 nm particle diameter was drop-casted on the carbon tracks printed on the substrate made of alumina ceramic. After evaporation, the electrode was carefully dipped in copper plating solution for a certain time to achieve a sufficient penetration of solution within the polystyrene spheres. The metal was then electrodeposited galvanostatically over the self-assembled colloidal crystal. Finally, the polystyrene template was dissolved in toluene to expose the porous structure of copper deposit. The morphology of porous structures was investigated using scanning electron microscopy. Electroanalytical properties of porous copper film electrodes were evaluated in amperometric detection of selected saccharides, namely glucose, fructose, sucrose, and galactose. Using hydrodynamic amperometry in stirred alkaline solution, a current response at +0.6 V vs. Ag/AgCl was recorded after addition of the selected saccharide. These saccharides could be quantified in two linear ranges (0.2-1.0 µmol L-1 and 4.0-100 µmol L-1) with detection limits of 0.1 µmol L-1 glucose, 0.03 µmol L-1 fructose, and 0.05 µmol L-1 sucrose or galactose. In addition, analytical performance of porous copper electrodes was ascertained and compared to that of copper film screen-printed carbon electrodes, prepared ex-situ by the galvanostatic deposition of metal in the plating solution. After calculating the current densities with respect to the geometric area of working electrodes, the porous electrodes exhibited much higher sensitivity to changes in concentration of analytes, presumably due to the larger surface of the porous copper deposit. In the future, they could be incorporated in detectors of flow injection systems due to their long-term mechanical stability.

Analytical Performance of Clay Paste Electrode and Graphene Paste Electrode-Comparative Study.

The analytical performance of the clay paste electrode and graphene paste electrode was compared using square wave voltammetry (SWV) and cyclic voltammetry (CV). The comparison was made on the basis of a paracetamol (PA) determination on both working electrodes. The influence of pH and SWV parameters was investigated. The linear concentration ranges were found to be 6.0 × 10-7-3.0 × 10-5 and 2.0 × 10-6-8.0 × 10-5 mol L-1 for clay paste electrode (ClPE) and graphene paste electrode (GrPE), respectively. The detection and quantification limits were calculated as 1.4 × 10-7 and 4.7 ×10-7 mol L-1 for ClPE and 3.7 × 10-7 and 1.2 × 10-6 mol L-1 for GrPE, respectively. Developed methods were successfully applied to pharmaceutical formulations analyses. Scanning electron microscopy and energy-dispersive X-ray spectroscopy were used to characterize ClPE and GrPE surfaces. Clay composition was examined with wavelength dispersive X-ray (WDXRF).

Voltammetric Quantification of Anti-Cancer Antibiotic Bleomycin Using an Electrochemically Pretreated and Decorated with Lead Nanoparticles Screen-Printed Sensor.

In this paper, we report a highly sensitive voltammetric sensor for the determination of the anti-cancer antibiotic bleomycin (BLM) based on a screen-printed carbon sensor that is electrochemically pretreated and decorated with lead nanoparticles in the sample solution (pSPCE/PbNPs). These sensor surface manipulations contribute to significant amplification of the analytical signal and improvement of its shape and repeatability. The effect of the electrochemical behavior of BLM on the pSPCE/PbNPs was examined by electrochemical strategies. CV, EIS, and XPS were used to compare the sensor surface modifications. The effects of the type and pH of the supporting electrolyte and the procedure parameters were optimized. The features of the proposed procedure include: (a) very low limits of detection and quantification (2.8 × 10-11 and 9.3 × 10-11 M, respectively), (b) linear ranges (1.0 × 10-10-2.0 × 10-9 M and 2.0 × 10-9-2.0 × 10-8 M, and (c) a high sensitivity of 0.32 µA/nM. The electrochemical sensor was successfully applied for the determination of BLM in wastewater and reference material of human urine samples.

A new voltammetric approach for the determination of ß-carotene in vegetables and pharmaceutical capsules using a gold electrode.

Significant improvements in the voltammetric determination of ß-carotene (BCA) have been achieved, mainly by the replacement of toxic dichloromethane with acetone and using non-mercury electrode. The respective procedure is based on anodic oxidation of BCA at a gold electrode in the disc configuration, when using square-wave voltammetry in pure acetone (99.8%) with 0.1 mol L-1 LiClO4 as the supporting electrolyte. The method comprises extraction of the analyte from the sample with acetone, thus avoiding the usually used highly toxic solvents. Analytically, it can be characterized by a linear range from 6.0 × 10-6 to 5.9 × 10-4 mol L-1 with regression equation Ipa = 0.0184c -0.1631 and correlation coefficient, R2 = 0.9998, limits of detection and quantification LOD = 1.6 × 10-6 mol L-1 and LOQ = 5.4 × 10-6 mol L-1, respectively; both being obtained at a potential step of 5 mV, with the pulse amplitude of 25 mV, and a frequency of 80 Hz. After optimization, the method was evaluated in series of analyses; namely, with two samples of vegetables and two pharmaceutical preparations (capsules), when the results could be compared to those of a reference spectrophotometric method. Due to a simple instrumentation, including sample preparation, the voltammetric method for the determination of BCA can be recommended as a quick screening assay in food and pharmaceutical analysis.

Rapid monitoring of fungicide fenhexamid residues in selected berries and wine grapes by square-wave voltammetry at carbon-based electrodes.

A completely new electroanalytical method for the determination of fenhexamid (FNX) residues in fruit samples has been developed. This method is based on anodic oxidation of fungicide in Britton-Robinson buffer (pH 4) containing 10% (v/v) methanol using square-wave voltammetry when five different carbon-based electrodes were tested. An electrochemical behaviour of FNX was studied on a glassy carbon electrode using cyclic voltammetry, while glassy carbon paste electrode was selected for analytical purposes. Linear range for FNX from 3.96 to 49.50 µmol L-1 characterized by coefficient of determination of 0.9964, sensitivity of 0.176 µA L µmol-1, and detection limit of 1.32 µmol L-1 were calculated. Results acquired from analyses of blueberries and wine grapes were compared to those obtained by a reference chromatographic method, and a satisfactory agreement has been reached. Finally, it seems that the present voltammetric approach could find its application in food quality control as screening assay.

Multichannel separation device with parallel electrochemical detection.

A device with four parallel channels was designed and manufactured by 3D printing in titanium. A simple experimental setup allowed splitting of the mobile phase in four parallel streams, such that a single sample could be analysed four times simultaneously. The four capillary channels were filled with a monolithic stationary phase, prepared using a zwitterionic functional monomer in combination with various dimethacrylate cross-linkers. The resulting stationary phases were applicable in both reversed-phase and hydrophilic-interaction retention mechanisms. The mobile-phase composition was optimized by means of a window diagram so as to obtain the highest possible resolution of dopamine precursors and metabolites on all columns. Miniaturized electrochemical detectors with carbon fibres as working electrodes and silver micro-wires as reference electrodes were integrated in the device at the end of each column. Experimental separations were successfully compared with those predicted by a three-parameter retention model. Finally, dopamine was determined in human urine to further confirm applicability of the developed device.

Electrochemical quantum dots-based magneto-immunoassay for detection of HE4 protein on metal film-modified screen-printed carbon electrodes.

A novel enzyme-free electrochemical immunosensor was developed for highly sensitive detection and quantification of human epididymis protein 4 (HE4) in human serum. For the first time, core/shell CdSe/ZnS quantum dots were conjugated with anti-HE4 IgG antibodies for subsequent sandwich-type immunosensing with superparamagnetic microparticles functionalized with anti-HE4 IgG antibodies, which allow rapid and efficient HE4 capture from the sample. Electrochemical detection of anti-HE4 IgG - HE4 - anti-HE4 IgGCdSe/ZnS immunocomplex was performed by recording the current response of Cd(II) ions, released from dissolved quantum dots at screen-printed carbon electrode (SPCE), modified with mercury or bismuth film. The linear range of the detection was from 20 pM to 40 nM with limit of detection of 12 pM using three times the standard deviation of blank criterion at mercury-film SPCE and from 100 pM to 2 nM with limit of detection of 89 pM at bismuth-film SPCE. Proposed electrochemical immunosensor meets the requirements for fast and sensitive quantification of HE4 biomarker in early stage of ovarian cancer and due to the proper sensitivity and specificity presents a promising alternative to enzyme-based probes used routinely in clinical diagnostics.

Recent Applications of Silver Amalgam Electrodes for Analysis of Pharmaceuticals and Vitamins.

The features and applications of silver amalgam electrodes in electroanalysis of pharmaceuticals and vitamins are summarized in this review. The state-of-the-art in the preparation and construction of solid silver amalgam electrodes for prolonged and userfriendly use is presented. Although not as widely spread as other electrode materials, silver amalgam possesses a unique and viable combination of favorable electrochemical properties, which are close to ideal mercury electrodes, non-toxic character, high durability and mechanical stability, if properly prepared. Its capability of conducting the redox processes at highly negative potentials is essential for the analysis of large number of bioactive organic compounds. The review features also overview information for each application in drug or vitamin analysis divided to the sections according to the construction variants of silver amalgam electrodes.

Monolithic capillary column with an integrated electrochemical detector.

The carbon fiber and silver microwire were used as working and pseudoreference electrode, respectively, and inserted into the ending of capillary to prepare monolithic capillary column with an integrated electrochemical detector. Prepared capillary devices offered stable and robust results with relative standard deviations of retention, resolution, and detection signal lower than 1.5, 5.5, and 5.0%, respectively. To further increase sensitivity of developed electrochemical microdetector, multiple pulse amperometry detection mode has been used. Optimized integrated device provided reliable chromatographic separation of mixture of neurotransmitters with calibration curve for dopamine linear from 0.5 to 20.0mgL-1 and an instrumental limit of detection as low as 24pg of injected dopamine. Finally, developed capillary column was applied to successful determination of dopamine in a human urine. By using both calibration curve and standard addition method, the dopamine level was determined to be 0.74±0.03mgL-1 and 0.71±0.02mgL-1, respectively. Triplicates of dopamine analysis provided relative standard deviations lower than 2.7% for intraday analyses, while interday relative standard deviations were lower than 3.6% for five consecutive days.

Determination of vitamin E in margarines and edible oils using square wave anodic stripping voltammetry with a glassy carbon paste electrode.

A new electroanalytical method for determination of vitamin E in the form of the total content of tocopherols present in margarines and edible oils has been developed. The method is based on extraction of these biologically active compounds into silicone oil, acting as lipophilic binder of glassy carbon paste electrode, with subsequent electrochemical detection by square wave anodic stripping voltammetry (SWASV) in 0.1M HNO3. The values of vitamin E contents were expressed as mass equivalent of α-tocopherol known as the most active form of this lipophilic vitamin. The linear ranges for α-tocopherol determination were 5×10-7-4×10-5 and 5×10-8-1×10-5molL-1 with the detection limits of 1×10-7 and 3.3×10-9molL-1 for 5 and 15min accumulation, respectively. The results have shown that SWASV with extraction step is very sensitive method for the determination of vitamin E, being comparable to reversed-phase high performance liquid chromatography chosen as reference method.

Improved electroanalytical characteristics for flumetralin determination in the presence of surface active compound.

ABSTRACT: The use of square wave voltammetry (SWV) and square wave adsorptive stripping voltammetry (SWAdSV) in conjunction with a cyclic renewable silver amalgam film electrode for the determination of flumetralin is presented. Poor separation of two overlapped reduction peaks is significantly improved when hexadecyltrimethylammonium bromide is used as a component of the supporting electrolyte solution (together with BR buffer pH 9.5). The SW technique parameters were investigated and found optimal as follows: frequency 50 Hz, amplitude 40 mV, and step potential 5 mV. Accumulation time and potential were studied to select the optimal conditions in adsorptive voltammetry. The analytical curve was linear for the flumetralin concentration range from 1.0 × 10-6 to 1.0 × 10-5 mol dm-3 and from 5.0 × 10-9 to 1.0 × 10-7 mol dm-3 for SWV and SWAdSV, respectively. Detection limit of 6.5 × 10-10 mol dm-3 was calculated for accumulation time 60 s at -0.2 V. The repeatability of the method was determined at a flumetralin concentration level equal to 5.0 × 10-9 mol dm-3 and expressed as %RSD = 5.0% (n = 6). The proposed method was applied and validated successfully by studying the recovery of herbicide content in spiked environmental samples.

Magnetic beads-based electrochemical immunosensor for monitoring allergenic food proteins.

Screen-printed platinum electrodes as transducer and magnetic beads as solid phase were combined to develop a particle-based electrochemical immunosensor for monitoring the serious food allergen ovalbumin. The standard arrangement of enzyme-linked immunosorbent assay became the basis for designing the immunosensor. A sandwich-type immunocomplex was formed between magnetic particles functionalized with specific anti-ovalbumin immunoglobulin G and captured ovalbumin molecules, and secondary anti-ovalbumin antibodies conjugated with the enzyme horseradish peroxidase were subsequently added as label tag. The electrochemical signal proportional to the enzymatic reaction of horseradish peroxidase during the reduction of hydrogen peroxide with thionine as electron mediator was measured by linear sweep voltammetry. The newly established method of ovalbumin detection exhibits high sensitivity suitable for quantification in the range of 11 to 222nM and a detection limit of 5nM. Magnetic beads-based assay format using external magnets for rapid and simple separation has been proven to be an excellent basis for electrochemical detection and quantification of food allergens in highly complex sample matrices.

Carbon paste electrodes modified with Bi(2)O(3) as sensors for the determination of Cd and Pb.

Carbon paste electrodes bulk-modified with Bi(2)O(3)were used for the determination of Cd(II) and Pb(II). The best composition was 1% (wt%) Bi(2)O(3) in the paste. The measurements were made by differential pulse voltammetry in the potential range from -1.2 V to -0.3 V. The peak potential of the reoxidation of Cd is -0.85 V, and of Pb -0.60 V vs. SCE. The lowest concentration that could be determined was 5 microg L(-1) of both metals (preconcentration time 240 s), the relative standard deviation was 3.5%-5.0% (four determinations). The correlation coefficient ( r(2)) of the calibration curves was 0.9966 (for Cd) and 0.9971 (for Pb). The Bi(2)O(3)-modified electrode could be used for the analysis of drinking water, mineral water and urine.