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Biomedical subjects

Géza Nagy

Publications and source records attributed to Géza Nagy.

12 recordsLinked to original sources

Periodically interrupted amperometry. A way of improving analytical performance of membrane coated electrodes.

Amperometry is a powerful voltammetric measuring method. Its application is specially advantageous when used in combination with a separation step or with some other sample treatment method providing selectivity. The selectivity is often achieved by coating the amperometric working electrode surface with a membrane of special character. Size exclusion membrane, immobilized enzyme containing reaction layer, protecting dialysis membrane, perm selective ion exchange film etc can be mentioned here. In conventional amperometry the measuring potential is continuously applied, therefore in case of membrane coated electrodes the electrode process depletes the diffusion layer. In this work the performance of a new periodically interrupted amperometric (PIA) measuring program has been investigated in case of glucose enzyme sensor. The measuring program allowing time for reloading the diffusion layer provided higher current and therefore improved sensitivity and lower limit of detection.

Biosensing Techniques↗

Electrochemical behavior of ferrocene in ionic liquid media.

Chemistry and applicability of ionic liquids (IL), - organic salts with low melting point - are in the focus of interest today. The ILs with melting point below room temperature are expected to be good solvents. Their applicability in organic synthetic work, in separation processes as well as in electrochemistry is very promising. In the work reported here the voltammetric behavior of ferrocene in 1-butyl-3-methylimidazolium hexafluorophosphate (BMIM+ PF6-) ionic liquid has been investigated. Conventional size and micro platinum and carbon electrodes were employed in CV and in chronoamperometric measurements. Karl Fischer method was used for the determination of water content of the solvent. Voltammetric measurements without addition of background electrolyte could be carried out in (BMIM+ PF6-) ionic liquid. A broad potential window could be used. Concentration dependence of the electrochemically determined diffusion coefficient of the ferrocene was observed.

Carbon↗

How to assess the limits of ion-selective electrodes: method for the determination of the ultimate span, response range, and selectivity coefficients of neutral carrier-based cation selective electrodes.

The span and range of an ion-selective electrode (ISE) has been identified by IUPAC as a potential or activity difference between the upper and lower detection limits of the electrode. Once the span is known, the ultimately attainable detection limit of the ISE can be calculated using its theoretical response slope. In this paper, we propose an original method for the determination of the ultimate span and response range of ISEs. The simple measurement of span is recommended to aid the fast screening of novel ionophores and help to focus optimization processes to the most promising candidates. The measurement of span is combined with a generally applicable procedure for the determination of the three seminal parameters of ISEs: the response slope, the ultimate selectivity coefficients, and detection limit. In the proposed procedure, following the span measurement, two subsequent exponential dilution experiments are completed in which the responses of the electrode for the primary and the interfering ions are tested using a solution of a discriminated ion and deionized water as diluting electrolytes in consecution. The advantages and the practical usefulness of the proposed methods and procedures are demonstrated through the evaluation of the performance characteristics of novel and well-characterized ionophore-based potassium and calcium sensors.

Calcium↗

[Effect of vitamin E supplementation on the vitamin content of lipoprotein in young men and women].

UNLABELLED: The effect of supplementary Vitamin E on the vitamin content of lipoproteins in young men and women. Inappropriate vitamin and trace element supplementation may facilitate the development of atherosclerosis. It is known that Vitamin E protects lipids from oxidative stress, while clinical signs of atherosclerosis appear later in women compared to men. AIMS: (1) The increase of vitamin E in plasma and plasma lipoproteins after 4 weeks of supplementation vitamin E was investigated, (2) furthermore it was tested whether a proportion shift occurs in alpha-tocopherol content of lipoproteins, (3) and checked for gender-related differences in plasma and plasma lipoprotein vitamin E levels before, during and after treatment, (4) plasma CRP levels as a marker of lipid peroxidation were also followed. METHODS: 5-5 young healthy men and women took part in the study, receiving 700 IU/day Vitamin E for one month. Each subject was studied before and at the end of treatment, and also one month after treatment. HDL and LDL-VLDL containing lipoproteins were separated. Vitamin E and hsCRP levels were measured (by HPLC and an immunoturbidimetric method, respectively). RESULTS: Vitamin E treatment induced in both genders an approximately threefold increase in vitamin E concentration in HDL-cholesterol (8.1 +/- 1.7 micromol/l vs. 22.5 +/- 7.5 micromol/l, p < 0.001), and a twofold increase in LDL-VLDL-cholesterol (22.0 +/- 3.7 micromol/l vs. 49.0 +/- 9.0 micromol/l, p < 0.001). Plasma and HDL vitamin E levels were higher in women than in men at the onset of treatment (6.8 +/- 0.96 micromol/l vs. 9.5 +/- 1.10 micromol/l), but during the treatment these gender-related differences disappeared. When plasma vitamin E concentration were considered 100% and the changes of the vitamin E concentrations of lipoproteins were calculated, it was found that supplementation with vitamin E in men increased the vitamin E concentration of LDL-VLDL cholesterol to a higher extent compared to women (LDL-VLDL % in men: 59.8 +/- 7.43%, in women: 49.3 +/- 7.41%, p < 0.05). All the observed changes regressed one month after cessation of supplementation. The level of hsCRP decreased during vitamin E treatment (1.07 +/- 0.9 mg/l vs. 0.2 +/- 0.14 mg/l, p < 0.001), and remained suppressed after the cessation of treatment (0.37 +/- 0.4, p < 0.01). CONCLUSIONS: These results support the hypothesis that women at young age are better protected against lipid-peroxidation as compared to men because of higher HDL vitamin E concentrations. Vitamin E supplementation in men eliminates this concentration difference between genders, and also increases LDL-VLDL vitamin E. In both genders high concentration of vitamin E in lipoproteins was associated with low hsCRP concentration.

Adult↗

Unexpected effect of charge density of the aromatic guests on the stability of calix[6]arene-phenol host-guest complexes.

The pi-pi interaction-based inclusion complexation of calix[6]arene hexasulfonate as host with neutral aromatic guest molecules was studied in aqueous media. To vary the electron density on the guest's aromatic rings, the phenol parent compound was functionalized in the para-position with different electron-withdrawing groups, such as NO2 and Cl, as well as H and CH3 groups. To study the interaction between calixarene and the guests, PL, DSC, and quantum-chemical methods were used. The results indicate 1:1 stoichiometry for all examined host-guest complexes. Although the enthalpy change predicts strong interaction between the host and the guest, the Gibbs free energy change of the complex formation is small, resulting in a relatively low complex stability. This property is due to the high and negative entropy change during the complex formation. Comparing the thermodynamic parameters observed on the series of the guests, we observed a decrease of the enthalpy change when the electron density on the guest's aromatic ring increased. However, the Gibbs free energy and therefore, the stability of the complexes increased when the enthalpy change lowered. These unexpected results are based on the enthalpy-entropy compensation effect and probably due to the quite different entropy change related to the high and low electron density on the aromatic rings of different guest molecules. Using molecular dynamic calculations, a redistribution of the electron density of calixarene rings, followed by the reordering of the solvent molecules, was identified as a background of this unexpected entropy change at molecular level.

Journal Article↗

Determination of diffusion coefficient in gel and in aqueous solutions using scanning electrochemical microscopy.

Diffusion coefficient of different species in different media is an important property needed in scientific research and practice. A method taking advantage on the special capability of scanning electrochemical microscopy (SECM) is described for the easy and accurate measurement of diffusion coefficient. The method is based on detecting the concentration-time transients with appropriate electrochemical microsensor positioned at the close vicinity of a miniature dose-source device. At a given time (ti), a small dose of the investigated species is introduced. The Deltatmax=(tcmax-ti) value and the distance (d=x+Deltaxn) between the source and the detector microelectrode are used for the calculation of D. While the original set distance (x) cannot be accurately measured in the micrometer scale, the tip travel distance (Deltaxn) of the microscope is well defined. Collecting a few Deltatmax-(x+Deltaxn) data pairs, a reliable value of the diffusion coefficient can be obtained. The procedure is simple, and no exact knowledge of the introduced dose is needed. Two ways of sample dose delivery were used: on the one hand, coulometric generation with current-controlled electric pulse using micro-disc electrode, and on the other one, pressure ejection of a nano-droplet from a glass micropipette. Diffusion coefficient of I2, H2O2, [Ru(NH3)6]Cl3 and K3[Fe(CN)6] were measured in solution and in agarose gel phases of different composition. The effect of polyelectrolyte ion exchangers on the diffusion of the investigated species was checked.

Diffusion↗

Optical biosensor for urea with improved response time.

An optical biosensor for urea measurements was developed. The operation of the sensor is based on the well-known urease enzyme-catalyzed hydrolysis of urea. The ammonium ions liberated in the reaction are detected with an ion selective optode membrane containing nonactin as ion selective ionophore and ETH 5294 chromoionophore in a thin (1 microm) plasticized poly(vinylchloride) film. The basic sensing element was home made of a microscope glass slide, a HeNe laser light source, photodiode light detector and light in coupling, de-coupling elements. The transducer membrane and the enzyme containing reaction layer were sandwich-cast with spin coating onto the surface of the sensing slide. The attenuation of the laser light propagating inside the glass wave-guide was used as signal for urea measurements. With this arrangement membranes provided good sensitivity (0.05 absorption unit when going from 0.1 to 1 mM urea) and short (16-20 s) response time. Taking advantage on the improved response time, flow injection urea measurements were made in the 0.01-2 mM concentration range. Thirty sample/hour analysis-rate, good peak-to-peak reproducibility (RSD=0.02) and recovery (95-104%) was achieved with buffer diluted urea solutions. Applications for the analysis of real samples are planned to do in the future.

Biosensing Techniques↗

Investigation of concentration profiles inside operating biocatalytic sensors with scanning electrochemical microscopy (SECM).

Scanning electrochemical microscopy (SECM) with amperometric or potentiometric measuring tips was used to investigate biocatalytic reactions inside the enzyme layer of a biosensor during its operation. The well known glucose oxidase catalyzed oxidation of glucose has been selected for the studies. Local, instantaneous concentration of dissolved oxygen and hydrogen peroxide was studied observing the amperometric current while miniaturized potentiometric tip served for local pH measurements. Liquid enzyme layer immobilized with Cellophane membrane or cross linked polyacrilamide gel membrane containing entrapped enzyme served for biocatalytic media in the SECM imaging. Local maximum of H(2)O(2) and minimum of O(2) profiles were found at approximately 200 microm far from the substrate/enzyme layer boundary. From the experimental findings guidelines to design well functioning biocatalytic sensors could be concluded. The concentration profiles obtained with SECM techniques were compared with the results of simple model calculations carried out with the method of finite changes. Most of earlier made SECM studies dealing with enzyme reactions imaged the electrolyte being in contact with the immobilized enzyme. The data in our investigation, however, were collected inside the working catalytic layer.

Biosensing Techniques↗

Amperometric microcells for alkaline phosphatase assay.

To develop simple electrochemical immunoassays, a screen printed amperometric microcell with graphite working and Ag/AgCl reference electrodes was tested for the determination of alkaline phosphatase enzyme (ALP) and anti-humanIgG conjugated ALP (alpha-hIgG-ALP) activity in 5-10 microl samples. To ensure reproducible, steady state conditions, the working electrode surface was coated with mass-transport controlling hydrogel layer. The kinetic response curves of the hydrogel coated electrodes were linear. In addition, the hydrogel layer reduced the nonspecific adsorption of the alpha-hIgG-ALP conjugate on the working electrode surface. The measurements were made in the range of 2 divided by 4000 mU ml(-1) enzyme activities using ascorbic acid 2-phosphate (AAP) as the enzyme substrate. AAP is commercially available, non-toxic and has excellent stability. The sensitivity of the determinations was about 71% of the sensitivity which could be achieved using p-aminophenylphosphate (PAPP), a not easily accessible and unstable enzyme substrate. The experimentally determined kinetic parameters of the ALP enzyme catalyzed reactions were the same with the bare and hydrogel layer coated electrodes.

Alkaline Phosphatase↗

Anisotropy decay study on the host-guest interaction of distally dialkylated calix[4]arenes with 1-chloro-4-(trifluoromethyl)benzene.

The 'host' properties of distally dialkylated calix[4]arenes and 4-tert-butylcalix[4]arenes in the presence of 1-chloro-4-(trifluoromethyl)benzene (BFT) were studied in chloroform solvent by intensity-independent spectrofluorometric method. The anisotropy decay experiments were found as suitable method to indicate the host-guest complex formation but it is unable to determine the strength of supramolecular interaction.

Benzene↗

Development and study of an amperometric biosensor for the in vitro measurement of low concentration of putrescine in blood.

An amperometric biosensor was developed for the in vitro determination of putrescine in blood samples because elevated level of putrescine in blood can be a diagnostic indicator of certain kinds of cancer. The electrochemical transducer consisted of a flat form, three electrode amperometric micro-cell fabricated with thin film photolithography on flexible Kapton substrate. An immobilized putrescine oxidase (PUO) layer provided the biocatalytic oxidation of the putrescine, while the generated hydrogen peroxide was detected on the platinum-working electrode. An electropolymerized poly(m-phenylenediamine) (pPDA) size-exclusion layer was used to protect the working electrode from fouling and to prevent signal generation by common electroactive interferents present in blood. The preparation of the biocatalytic enzyme- and outer protective layers was optimized for improved sensitivity and response time. A detection limit of 50 nM was achieved in pH-adjusted whole blood samples, which is below pathological levels.

Biosensing Techniques↗

Opto-electrochemical planar wave-guide sensor for copper (II) ion.

An opto-electrochemical sensor for copper (II) ion was developed. The sensor consists of a planar conductive indium-tin-oxide (ITO) glass support, which was coated by a polymeric copper (II) sensitive membrane. The sensing membrane is made of plasticized polyurethane matrix containing Zincon colorimetric reagent immobilized as ion pair with tetraoctylammonium ion. The instrumentation used commonly for wave-guide sensor development was extended with a potentiostat that made possible the control of the electric potential of the conductive surface. In this way the transport rate of ionic species through the membrane-sample interface could be influenced. In the presence of copper (II) ions the color of the membrane turned from red to blue, which was monitored optically. By applying positive potential to the conductive surface the direction of the ion diffusion at the membrane/sample interface was changed. As a result the sensing layer was regenerated within 2 minutes and was ready for further measurement. The sensor measured Cu(2+) ion in a concentration range of 1-200 microM.

Azo Compounds↗