Biomedical subjects

Shin-ichi Wakida

Publications and source records attributed to Shin-ichi Wakida.

11 recordsLinked to original sources

PubMed2006

On-chip micellar electrokinetic chromatographic separation of phenolic chemicals in waters.

This paper describes on-chip micellar electrokinetic chromatography (MEKC) separation of bisphenol A and 3 kinds of alkylphenols, which have been recently recognized as endocrine disrupting chemicals for fish by the Japanese government, using microchip capillary electrophoresis with UV detection. We successfully obtained high-speed separation of the phenolic chemicals within 15 s as optimizing in microfluidic controls and MEKC separation conditions. We obtained fairly good linearity with correlation coefficient of over 0.98 from 0 to 50 mg/l phenolic chemicals except for 4-nonylphenol, which sample is the mixture of many geometrical isomers (r = 0.86). The values of the relative standard deviation for peak height in 50 mg/l phenolic chemicals were less than 8% except for bisphenol A (11.0%). The limits of detection obtained at a signal-to-noise ratio of 3 were from 5.6 to 20.0 mg/l. To realize on-site monitoring, we described strategy for on-chip MEKC analysis of the phenolic chemicals in waters using a portable analyzer based on microfluidic devices.

Chromatography, Micellar Electrokinetic Capillary↗

PubMed2006

High-throughput nitric oxide assay in biological fluids using microchip capillary electrophoresis.

In order to develop a high-throughput screening method for the nitrogen monoxide metabolites, nitrite and nitrate, in biological fluids, we have investigated the simultaneous determination of these metabolites using microchip capillary electrophoresis (MCE). In this study, the control of applied voltage to obtain higher sensitivity by increasing the sample injection volume was investigated. Also, the improvement of reproducibility by correcting the injection volume using the internal standard was investigated. By increasing the sample volume, the limits of detection achieved for nitrite and nitrate were 24 and 12 microM, respectively. Because we used a 10-fold diluted sample when detecting nitrite and nitrate in human serum, it was necessary to increase the sensitivity by a factor of 10-50. The run-to-run and day-to-day relative standard deviations achieved were improved to less than 10% by using an internal standard to correct the injection volume. Moreover, we obtained successful separation of nitrite and nitrate in spiked human serum within 6.5 s under optimum analytical conditions. As a result, although it is necessary to obtain greater sensitivity, it was concluded that determination of the amount of NO metabolites in biological fluids using MCE is possible.

Calibration↗

PubMed2005

Development of an analytical method using microchip capillary electrophoresis for the measurement of fluorescein-labeled salivary components in response to exercise stress.

We have developed an analytical method using microchip capillary electrophoresis (microchip CE) for the high-speed separation of fluorescein-labeled salivary components in response to exercise stress. Optimal separation was obtained using a borate buffer at pH 9.5 containing 10 mM beta-cyclodextrin and 1.0% (w/v) methylcellulose. To minimize individual differences in human saliva, such as viscosity, conductivity, and contaminants, the concentration of methylcellulose in the analytical conditions played a key factor. The optimized separation conditions produced identical electropherograms successfully despite of the use of different microchips made from quartz glass or poly-methylmethacrylate (PMMA). In addition, a practical application of bicycle ergometer stress was performed. Some components in human saliva showed a marked decrease after exercise stress.

Electrophoresis, Microchip↗

PubMed2005

Determination of ammonium in river water and sewage samples by capillary zone electrophoresis with direct UV detection.

We developed capillary zone electrophoresis (CZE) with direct UV detection for determination of ammonium in environmental water samples. Ammonium in the samples was partly converted into ammonia in the alkaline background electrolyte (BGE) during migration and was detected by molecular absorption of ammonia at 190 nm in approximately 7 min. The limit of detection (LOD) for ammonium was 0.24 mg/l (as nitrogen) at a signal-to-noise ratio of three. The respective values of the relative standard deviation (RSD) of peak area, peak height, and migration time for ammonium were 2.1, 1.8, and 0.46%. Major alkali and alkaline earth metal ions coexisting in the samples did not interfere with ammonium determination by the proposed method. The proposed method determined ammonium in surface water and sewage samples. The results were compared to those obtained using ion chromatography (IC).

Calibration↗

PubMed2005

Thickness measurements on transparent substrates based on reflection ellipsometry. I. Optical effects of high-refractive-index additional layers.

Theoretical studies were conducted for thickness measurements using transparent substrates on the external and internal reflection configurations. For three-phase systems consisting of ambient, film, and substrate, the refractive index of the substrate could be optimized to obtain the high sensitivity of an ellipsometric quantity delta to the film thickness and the small susceptibility of delta to errors in the incident angle. It was shown that the combination of an ordinary glass substrate and an additional dielectric layer with an appropriate layer thickness works as a synthetic high-index single substrate (SHIS). The optical effect of the combination was approximately described by use of the effective refractive index of SHIS. A method to select the refractive index of the additional layer was also given.

Journal Article↗

PubMed2005

Two-dimensional thickness measurements based on internal reflection ellipsometry.

An imaging ellipsometer technique on internal reflection geometry that can measure the thickness distribution of a thin film possessing an assumed refractive index is described. Because a prism is used for the internal reflection geometry, it was theoretically predicted that angular derivation from the normal incidence on the prism surface affects only the psi value by a factor of 0.97 at maximum. Measurements were carried out for an optical system of silica substrate-TiO2 layer-silica layer-protein film-air, with a thin-film array of dried protein as the sample film. Thickness of the protein films was two-dimensionally estimated only from the measured map of the delta value by use of the simulated relationship between the thickness and the delta value. The thickness map obtained was coincident on the whole with the results according to a mechanical scanning. The detection limit was approximately +/- 0.2 nm. These findings validate the optical effect of a high-index additional layer to improve the sensitivity and precision of thickness measurements of the sample film on transparent substrates.

Coated Materials, Biocompatible↗

PubMed2003

Development of a novel running buffer for the simultaneous determination of nitrate and nitrite in human serum by capillary zone electrophoresis.

In order to improve NO2- peak height and obtain a convenient buffer system for the assay of nitrogen monooxide metabolites, we developed a novel running buffer for the simultaneous determination of nitrite and nitrate in human serum by capillary electrophoresis. The addition of cetyltrimethylammonium chloride to the running buffer resulted in high-speed separation using reverse electroosmotic flow. Highly sensitive determination was also achieved using stacking with 10-fold diluted sample solutions. The samples were injected hydrodynamically for 100 s into a 50 cm x 75 microm I.D. capillary. The separation voltage was 10 kV (negative polarity). UV detection was performed at 214 nm. We obtained complete separation of nitrite and nitrate in deproteinized human serum within 6 min with optimum analytical conditions. Linear calibration curves for nitrite and nitrate for both peak height and peak area were obtained with standard addition method. The limits of detection obtained at a signal-to-noise ratio of 3 for nitrite and nitrate were 4.1 and 2.0 microM, while the values of relative standard deviation of peak height were 2.4 and 2.6%, respectively.

Buffers↗

PubMed2003

Separation and on-line concentration of bisphenol A and alkylphenols by micellar electrokinetic chromatography with anionic surfactant.

Separation and on-line concentration of bisphenol A and three alkylphenols were investigated by micellar electrokinetic chromatography with the anionic surfactant, sodium dodecyl sulfate. The separation conditions were optimized by the simultaneous addition of the organic solvent and cyclodextrin to the running solution. The separation of hydrophobic analytes and 4-nonylphenol isomers was improved by the addition of 10% methanol and 5 mM beta-cyclodextrin to the running solution. When the sweeping with the running solution was used as the on-line concentration procedure, 69-, 48-, 55- and 41-fold increases in detection sensitivity were obtained for bisphenol A, 4-tert.-butylphenol and 4-(1,1,3,3-tetramethylbutyl)phenol, and the second peak of 4-nonylphenol isomers, respectively. The detection limits were 0.0071, 0.0065, 0.021 and 0.055 mg/l, respectively. These results were better than those with the cationic surfactant, tetradecyltrimethylammonium bromide.

Benzhydryl Compounds↗

PubMed2003

Stabilizing effect of artificial stabilizers for binding activity of QCM immunosensors.

We compare stabilizing effects of artificial and natural stabilizers for maintenance of binding activity of immobilized antibody on the gold surface of quartz crystal microbalance (QCM). Binding activity between immobilizing antibody and specific antigen was measured and considered as a calculated frequency shift with QCM-immunosensor after temperature increasing and long-term storage. According to the experimental results, one stabilizer among eight samples shows the best stabilizing effect for QCM-immunosensor.

Adsorption↗

PubMed2002

Separation and on-line concentration of bisphenol A and alkylphenols by micellar electrokinetic chromatography with cationic surfactant.

The separation and on-line concentration of bisphenol A and three alkylphenols were investigated by micellar electrokinetic chromatography with cationic surfactant. Tetradecyltrimethylammonium bromide was used as surfactant and the separation conditions were optimized by the addition of the organic solvents and cyclodextrins to the running solution. The separation of hydrophobic analytes and 4-nonylphenol isomers was improved by the addition of 20% acetonitrile and 20 mM beta-cyclodextrin to the running solution. When the sweeping with the running solution used as the on-line concentration procedure, 56-, 67- and 29-fold increase in detection sensitivity of bisphenol A, 4-tert.-butylphenol and 4-(1,1,3,3-tetramethylbutyl)phenol, respectively. The detection limits were 0.030, 0.098 and 0.159 mg/l, respectively.

Benzhydryl Compounds↗

PubMed2002

Determination of nitrite and nitrate in a proposed certified reference material for nutrients in seawater by capillary zone electrophoresis with artificial seawater as the background electrolyte using transient isotachophoresis.

We describe a combination of selected ions as a terminating ion which is useful for transient isotachophoresis (ITP) in capillary zone electrophoresis (CZE) for the determination of nitrite and nitrate in seawater. In addition to 150 mM sulfate as the principal terminating ion, 10 mM bromate was added to a sample solution as the additional terminating ion. Artificial seawater containing 3 mM cetyltrimethylammonium chloride (CTAC) was adopted as a background electrolyte (BGE). The limits of detection (LODs) for nitrite and nitrate were 2.2 and 1.0 microg/L (as nitrogen), respectively. The LODs were obtained at a signal to noise ratio (S/N) of 3. The values of the relative standard deviation (RSD) of peak area for these ions were 1.9 and 1.4%. The RSDs of peak height were 1.7 and 1.9%, the RSDs of migration time 0.11%. The proposed method was applied to the determination of nitrite and nitrate in a proposed certified reference material for nutrients in seawater, MOOS-1, distributed by the National Research Council of Canada (NRC). The results almost agreed with the assigned tolerance interval.

Calibration↗