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Dive into the research topics where Kenji Sueyoshi is active.

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Featured researches published by Kenji Sueyoshi.


Analytical Chemistry | 2008

On-Line Sample Preconcentration and Separation Technique Based on Transient Trapping in Microchip Micellar Electrokinetic Chromatography

Kenji Sueyoshi; Fumihiko Kitagawa; Koji Otsuka

This paper describes a novel on-line sample preconcentration and separation technique named transient trapping (tr-trapping), which improves the efficiencies of separation and concentration by using a partially injected short micellar plug in microchip electrophoresis. Although a longer separation length often provides a better resolution of complexed or closely migrating analytes, our proposed theoretical model indicated that a trap-and-release mechanism enables a short micellar zone, which was partially injected into the separation channel, to work as an effective concentration and separation field. Application of the tr-trapping technique to microchip micellar electrokinetic chromatography (MCMEKC) was performed on a newly fabricated 5-way-cross microchip by using sodium dodecyl sulfate and rhodamine dyes as test micelle and analytes, respectively. When the injection times of micelle (t(inj),M) and sample solution (t(inj),S) were 1.0 and 2.0 s, respectively, both the preconcentration and separation of the dyes were completely finished within only 3.0 s. At t(inj),S of 8.0 s, a 393-fold improvement of the detectability was achieved in comparison with conventional MCMEKC. The resolution obtained with tr-trapping-MCMEKC was also better than that with conventional MCMEKC in spite of the 160-fold shorter length of the injected micellar zone at t(inj),M of 1.0 s. These results clearly demonstrated that the tr-trapping technique in MCMEKC provides a rapid, high-resolution and detectability analysis even in the short separation channel on the microchips.


Journal of Chromatography A | 2012

Highly sensitive chiral analysis in capillary electrophoresis with large-volume sample stacking with an electroosmotic flow pump

Takayuki Kawai; Hiroshi Koino; Kenji Sueyoshi; Fumihiko Kitagawa; Koji Otsuka

To improve the sensitivity in chiral analysis by capillary electrophoresis without loss of optical resolution, application of large-volume sample stacking with an electroosmotic flow pump (LVSEP) was investigated. Effects of the addition of cyclodextrin (CD) into a running solution on the LVSEP preconcentration was theoretically studied, where the preconcentration efficiency and effective separation length would be slightly increased if the effective electrophoretic velocity (v(ep,eff,BGS)) of the analytes was decreased by interacting with CD. In LVSEP-CD-modified capillary zone electrophoresis (CDCZE) and LVSEP-CD electrokinetic chromatography with reduced v(ep,eff,BGS), up to 1000-fold sensitivity increases were achieved with almost no loss of resolution. In LVSEP-CD-modified micellar electrokinetic chromatography of amino acids with increased v(ep,eff,BGS), a 1300-fold sensitivity increase was achieved without much loss of resolution, indicating the versatile applicability of LVSEP to many separation modes. An enantio-excess (EE) assay was also carried out in LVSEP-CDCZE, resulting in successful analyses of up to 99.6% EE. Finally, we analyzed ibuprofen in urine by desalting with a C₁₈ solid-phase extraction column. As a typical result, 250ppb ibuprofen was well concentrated and optically resolved with 84.0-86.6% recovery in LVSEP-CDCZE, indicating the applicability of LVSEP to real samples containing a large amount of unnecessary background salts.


Journal of Chromatography A | 2012

Highly sensitive oligosaccharide analysis in capillary electrophoresis using large-volume sample stacking with an electroosmotic flow pump.

Takayuki Kawai; Masato Watanabe; Kenji Sueyoshi; Fumihiko Kitagawa; Koji Otsuka

To obtain high sensitivity in capillary electrophoresis of oligosaccharide without reducing the high resolution with an easy experimental procedure, large-volume sample stacking with an electroosmotic flow pump (LVSEP) was investigated. As a fundamental study, effect of the conductivity of a sample solution in LVSEP was examined. It was revealed that LVSEP was successfully carried out even in using a sample solution with the ionic strength of 150 μM and the conductivity ratio of 20, indicating a good applicability of LVSEP to the analysis of real samples containing salts. When glucose oligomer was analyzed as a model sample in LVSEP-capillary zone electrophoresis (CZE), all peaks were well resolved with decreasing only 5% of the peak-to-peak distance, which suggested 95% of the whole capillary could be used for the effective separation. In the analysis of maltoheptaose, a good calibration line with correlation coefficient of 0.9995 was obtained. The limit of detection was estimated as 2 pM, which was 500-fold lower than that in the conventional CZE. N-linked glycans released from three glycoproteins, bovine ribonuclease B, bovine fetuin, and human α(1)-acid glycoprotein were also analyzed by LVSEP-CZE. By the sample purification with a gel filtration column, further sample dilution to reduce the sample conductivity for LVSEP was not needed. All glycan samples were well concentrated and separated with up to a 770-fold sensitivity increase. The run-to-run repeatabilities of the migration time, peak height, and peak area were good with relative standard deviations of 0.1-1.3%, 1.2-1.7%, and 2.8-4.9%, respectively.


Journal of Pharmaceutical and Biomedical Analysis | 2014

Effective determination of a pharmaceutical, sulpiride, in river water by online SPE-LC-MS using a molecularly imprinted polymer as a preconcentration medium.

Takuya Kubo; Kenta Kuroda; Yuichi Tominaga; Toyohiro Naito; Kenji Sueyoshi; Ken Hosoya; Koji Otsuka

We report an effective and a quantitative analysis method for one of pharmaceuticals, sulpiride, in river water by online solid phase extraction (SPE) connected with liquid chromatography-mass spectrometry (LC-MS) using a molecularly imprinted polymer as a preconcentration medium. The polymer prepared with a pseudo template molecule showed the selective retention ability based on the interval recognition of functional groups in sulpiride. Also, the imprinted polymer provided an effective concentration of a trace level of sulpiride in offline SPE with dual washing processes using water and acetonitrile, although another imprinted polymer prepared by an authentic method using sulpiride and methacrylic acid as a template and a functional monomer, respectively, showed the selective adsorption only in organic solvents. Furthermore, we employed the imprinted polymer as the preconcentration column of online SPE-LC-MS and the results supposed that the proposed system allowed the quantitative analysis of sulpiride with high sensitivity and recovery (10ng/L at 96%). Additionally, the determination of sulpiride in real river water without an additional spiking was effectively achieved by the system.


Journal of Pharmaceutical and Biomedical Analysis | 2010

One-step preparation of amino-PEG modified poly(methyl methacrylate) microchips for electrophoretic separation of biomolecules

Fumihiko Kitagawa; Kei Kubota; Kenji Sueyoshi; Koji Otsuka

A simple method for a chemical surface modification of poly(methyl methacrylate) (PMMA) microchips with amino-poly(ethyleneglycol) (PEG-NH(2)) by nucleophilic addition-elimination reaction was developed to improve the separation efficiency and analytical reproducibility in a microchip electrophoresis (MCE) analysis of biomolecules such as proteins and enantiomers. In our procedure, the PEG chains were robustly immobilized only by introducing an aqueous solution of PEG-NH(2) into the PMMA microchannel. The electroosmotic mobilities on the modified chips remained almost constant during 35 days with 37 runs without any recoating. The PEG-NH(2) modified chip provided a fast, reproducible, efficient MCE separation of proteins with a wide variety of isoelectric points within 15s. Furthermore, the application of the modified chip to affinity electrophoresis using bovine serum albumin gave a good chiral separation of amino acids.


Journal of Chromatography A | 2012

Electrophoretic analysis of cations using large-volume sample stacking with an electroosmotic flow pump using capillaries coated with neutral and cationic polymers

Takayuki Kawai; Jun Ito; Kenji Sueyoshi; Fumihiko Kitagawa; Koji Otsuka

To realize the high-performance and simple-operation analysis of cationic compounds in capillary electrophoresis, we investigated large-volume sample stacking with an electroosmotic flow pump (LVSEP) using capillaries with hydrophilic and weakly cationic inner surface. Three capillary modification methods were employed: thermally passivated physical coating with polymer mixture of poly(vinyl alcohol) and poly(allylamine); covalent modification with random copolymer of acryl amide and 3-(methacryloylamino)propyltrimethylammonium chloride; easily preparable physical coating with dimethyldioctadecylammonium bromide and polyoxyethylene stearate. In these capillaries, the electroosmotic flow (EOF) was well suppressed in the high ionic strength (I) electrolyte under the acidic and basic pH, whereas the EOF was enhanced in the low I electrolyte, indicating a suitable EOF property for the rapid LVSEP and following separation. In the LVSEP-capillary zone electrophoresis (CZE) analyses of benzylamine and 1-naphthylethylamine, up to 550-fold sensitivity increases were successfully obtained in the three capillaries without significantly reducing the repeatability and resolution. LVSEP-cyclodextrin-modified CZE of chlorpheniramine and brompheniramine was also carried out, resulting in up to 380-fold sensitivity enhancement with keeping the baseline separation for the enantiomers. Finally, we performed the LVSEP-CZE analysis of basic proteins, where up to 100-fold sensitivity increases were achieved, but a peak broadening was observed due to the sample adsorption in the low I sample matrix.


Science and Technology of Advanced Materials | 2006

One-step immobilization of cationic polymer onto a poly(methyl methacrylate) microchip for high-performance electrophoretic analysis of proteins

Fumihiko Kitagawa; Kei Kubota; Kenji Sueyoshi; Koji Otsuka

Abstract One-step covalent immobilization of poly(ethyleneimine) (PEI) onto poly(methyl methacrylate) (PMMA) substrates was investigated to achieve an efficient separation of basic proteins in microchip electrophoresis (MCE). The PEI-treated PMMA chip showed the anodic electroosmotic flow and its rate was almost kept stable during 32 days with over 50 runs. This longer stability of the prepared microchip indicated that the loss of PEI was successfully suppressed by the immobilization through the covalent bond. Furthermore, the PEI modification onto the PMMA chip could apparently reduce the surface adsorption of cationic proteins. In the MCE analysis on the PEI-modified microchip, two proteins were successfully separated within 30 s only utilizing a separation length of 5 mm. While the migration time of the protein gradually increased during only four consecutive runs on an untreated PMMA chip, reproducible analyses were attained by using the PEI immobilized microchip. These results demonstrated that Coulombic repulsion force generated between cationic PEI and basic proteins could avoid the irreversible adsorption of the analytes onto the PMMA surface, which provided a high-performance analysis medium for biogenic compounds.


Electrophoresis | 2011

Hydrophobic labeling of amino acids: Transient trapping–capillary/microchip electrophoresis

Kenji Sueyoshi; Kota Hashiba; Takayuki Kawai; Fumihiko Kitagawa; Koji Otsuka

Transient trapping (tr‐trapping) was developed as one of the on‐line sample preconcentration techniques to improve a low concentration‐sensitivity in microchip electrophoresis (MCE), providing highly effective preconcentration and separation based on the trap‐and‐release mechanism. However, a poor performance to hydrophilic analytes limited the applicability of tr‐trapping. To overcome this drawback, tr‐trapping was combined with a sample labeling using a hydrophobic reagent in CE. Three commercially available fluorescent dyes, fluorescein isothiocyanate, succinimidyl esters of Alexa Fluor 488 and BODIPY FL‐X, were tested as derivatization reagents to increase the hydrophobicity of amino acids (AAs) that were undetectable due to no fluorescence/UV‐absorbance. As a result, it was confirmed that BODIPY labeling allowed various AAs to be analyzed in tr‐trapping–micellar electrokinetic chromatography (tr‐trapping–MEKC) by the increase in the hydrophobicity. In tr‐trapping–MEKC, both the improvement of the resolution and 106–125‐fold enhancements of the detectability of labeled AAs were achieved relative to the conventional capillary zone electrophoresis. The limit of detection of labeled phenylalanine was improved from 800 to 5 pM by applying tr‐trapping–MEKC. In tr‐trapping–microchip MEKC, furthermore, an 80–160‐fold enhancement of the peak intensity and a baseline separation was also achieved within 30 s. These results clearly demonstrate that the tr‐trapping technique with hydrophobic labeling will make CE/MCE more sensitive for various analytes.


Analytical Biochemistry | 2013

Capillary-based enzyme-linked immunosorbent assay for highly sensitive detection of thrombin-cleaved osteopontin in plasma

Shun-ichi Funano; Terence G. Henares; Mie Kurata; Kenji Sueyoshi; Tatsuro Endo; Hideaki Hisamoto

In this study, a highly sensitive capillary-based enzyme-linked immunosorbent assay (ELISA) has been developed for the analysis of picomolar levels of thrombin-cleaved osteopontin (trOPN), a potential biomarker for ischemic stroke, in human plasma. Using a square capillary coated with 8.5 μg/ml anti-human trOPN capture antibody for ELISA, the linear range obtained was 2 to 16 pM trOPN antigen. This concentration range was in the detection window of trOPN antigen in plasma samples. Compared with the conventional microplate-based ELISA, the current capillary technique significantly reduced the amounts of reagent from milliliter to microliter, reduced the analysis time from 8 to 3 h, and had a better sensitivity and detection limit performance from approximately 50 pM down to 2 pM of trOPN antigen. These results indicate that this capillary-based immunoassay is a potential tool for biomarker detection and may be useful in clinical trials and medical diagnostic applications.


Electrophoresis | 2013

Toward 10 000-fold sensitivity improvement of oligosaccharides in capillary electrophoresis using large-volume sample stacking with an electroosmotic flow pump combined with field-amplified sample injection

Takayuki Kawai; Masumi Ueda; Yudai Fukushima; Kenji Sueyoshi; Fumihiko Kitagawa; Koji Otsuka

A combination of two online sample concentration techniques, large‐volume sample stacking with an electroosmotic flow pump (LVSEP) and field‐amplified sample injection (FASI), was investigated in CE to achieve highly sensitive oligosaccharide analysis. In CE with LVSEP‐FASI, analytes injected throughout the capillary were concentrated on the basis of LVSEP, followed by an electrokinetic introduction of concentrated analytes from the inlet vial by the FASI mechanism. After switching the inlet vial solution from the sample to running buffer, the concentrated analytes were then separated by CZE. In the present LVSEP‐FASI‐CZE, pressure was applied to the capillary inlet until the inlet vial solution was exchanged. The applied pressure generated a counterflow against the EOF. It kept the stacked sample zone within the capillary, minimizing loss of concentrated analytes. Fluorescein was first analyzed by LVSEP‐FASI‐CZE to optimize preconcentration condition. Up to 110 000‐fold sensitivity increase was obtained with 200 μL of sample, compared to normal CZE with sample injection of 0.3 psi for 3 s (ca. 1.7 nL). From the results, the pressure application improved the efficiency of the FASI‐mode concentration significantly at total concentration time longer than 10 min. In the analysis of maltoheptaose, a 10 000‐fold sensitivity increase was achieved, which is the highest concentration efficiency ever reported in CE of oligosaccharides. The relative standard deviations of the detection time and peak height were 2.4 and 11%, respectively. In the analysis of glucose oligomer, up to 8600‐fold sensitivity increases were achieved without reducing the separation performance of conventional CZE.

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Hideaki Hisamoto

Osaka Prefecture University

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Tatsuro Endo

Osaka Prefecture University

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Shoma Aki

Osaka Prefecture University

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Kenichi Maeno

Osaka Prefecture University

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Terence G. Henares

Osaka Prefecture University

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