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Application of a phase-separation multiphase flow as an eluent to a HPLC system using a core-shell silica-particle packed column

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JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES 2025年

作者： Kimura, Aoi Iharada, Takeshi tsukagoshi, kazuhiko Doshisha Univ Grad Sch Sci & Engn Appl Chem Kyotanabe 6100321 Japan

This study investigates the use of phase-separation multiphase flow (PSMF) as an eluent in a high-performance liquid chromatography (HPLC) system with a core-shell silica-particle packed column. Initially, twenty-two kinds of mixed solution, comprising combinations of water-acetonitrile as well as water-acetonitrile-ethyl acetate, were tested as an eluent at 20 degrees C. Two analytes as models, 2,6-naphthalenedisulfonic acid (2,6-NDS) and 1-naphthol (1-NA), were introduced into the system. In normal-phase mode with organic solvent-rich eluents, 1-NA was detected before 2,6-NDS. Conversely, with water-rich eluents in reverse-phase mode, 2,6-NDS was detected prior to 1-NA. Subsequently, ternary mixed solutions were tested at 0 degrees C. Seven of these solutions featured two-phase separation mixtures, forming a PSMF within the separation column at this lower temperature. Separation of the analyte mixture was observed with organic solvent-rich eluents at 20 degrees C (normal-phase mode) as well as 0 degrees C (phase-separation mode), 1-NA being detected before 2,6-NDS. Notably, separation efficiency improved at 0 degrees C compared to 20 degrees C. For water-rich eluents, separation occurred at both temperatures, with 2,6-NDS detected before 1-NA in reverse-phase mode at 20 degrees C and in phase-separation mode at 0 degrees C, where also the phase-separation mode improved separation efficiency than the reverse-phase mode. This information shed light on the phase-separation mode of HPLC possessing a core-shell silica-particle packed column.

关键词： Core-shell silica-particle packed column HPLC phase-separation mode phase-separation multiphase flow

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Effects of Porous Structure of Separation Column on Chromatograms Obtained by Phase-Separation Mode in HPLC

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CHROMATOGRAPHIA 2025年第3期88卷 215-224页

作者： Katayama, Tetsuro Iharada, Takeshi Obata, Yuki Sakamaki, Hiroshi tsukagoshi, kazuhiko Doshisha Univ Grad Sch Sci & Engn Appl Chem Kyotanabe 6100321 Japan Chem Evaluat & Res Inst 1600 ShimotakanoSugito Machi Saitama 3450043 Japan

A novel HPLC system utilizing a phase-separation multiphase flow as the eluent has been developed, referred to as phase-separation mode. This research explores the influence of the porous structure in an octadecyl-modified silica (ODS) column (with a pore diameter of 12 nm) on chromatographic outcomes under the phase-separation mode in HPLC. The chromatograms obtained from the porous ODS column were compared to those generated with a non-porous ODS column. In preliminary experiments, twenty-four mixed solutions, comprising combinations of water/acetonitrile/ethyl acetate and water/acetonitrile, were introduced as eluents at a column temperature of 20 degrees C. A model mixture of 2,6-naphthalenedisulfonic acid (2,6-NDS) and 1-naphthol (1-NA) was injected into the system, with separation achieved in most solutions except for some highly organic solvent-rich solutions where 2,6-NDS eluted faster than 1-NA, indicating reverse-phase mode operation. Subsequently, the separation of the model mixture was assessed at 0 degrees C, and four specific ternary mixtures were analyzed in detail at both 20 degrees C and 0 degrees C. These ternary mixtures, defined by their volume ratios, exhibited a two-phase separation, establishing a phase-separation multiphase flow. Consequently, the solution flow was homogeneous at 20 degrees C and heterogeneous at 0 degrees C. For instance, solutions with water/acetonitrile/ethyl acetate ratios of 20:60:20 (organic solvent-rich) and 70:23:7 (water-rich) were introduced as eluents at both 20 degrees C and 0 degrees C. At 0 degrees C in the organic solvent-rich eluent, 1-NA eluted faster than 2,6-NDS, characteristic of the phase-separation mode. In contrast, the water-rich eluent resulted in faster elution of 2,6-NDS at both temperatures. The porous ODS column displayed improved separation efficiency at 0 degrees C compared to the non-porous column, which can be attributed to the porous effect under phase-separation conditions.

关键词： Phase-separation multiphase flow Phase-separation mode Porous ODS column HPLC

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Reactive Oxygen Species (ROS) Generated on the Surface of (100)-Plane Grain-Oriented Copper Thin-Film

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Materials Sciences and Applications 2025年第1期16卷 27-45页

作者： Ken Hirota Taika Maeda kazuhiko tsukagoshi Yurika Taniguchi Hiroshi Kawakami Takashi Ozawa Masahiko Wada Research Center of Bio-Micro-Fluidic Science Doshisha University Kyoto Japan Department of Chemical Engineering and Materials Science Faculty of Science & Engineering Doshisha University Kyoto Japan Graduate School of Engineering Osaka Metropolitan University Osaka Japan Japan Copper Development Association Tokyo Japan

This work aims to study the dependence of the antibacterial activity on the crystal plane of Cu. The generation of reactive oxygen species (ROS) on the thin film of Cu with grains oriented in the plane (100) was evaluated by chemiluminescence (CL). The authors proposed the generation mechanism of these three ROS on the outermost surface consisting of Cu2O thin film, CuO layer and bulk Cu.

关键词： Copper Crystal Planes Antibacterial Activity Reactive Oxygen Species Chemiluminescence

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Fundamental Research and Application of the Specific Fluidic Behavior of Mixed Solvents in a Microspace

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ANALYTICAL SCIENCES 2014年第1期30卷 65-73页

作者： tsukagoshi, kazuhiko Doshisha Univ Dept Chem Engn & Mat Sci Kyoto 6100321 Japan Doshisha Univ Tube Radial Distribut Phenomenon Res Ctr Kyoto 6100321 Japan

The author herein reviews a specific microfluidic behavior exhibited by mixed-solvent solutions in a microspace, coined as the tube radial distribution phenomenon (TRDP). The specific fluidic behavior was observed in the following solution systems: ternary water-hydrophilic/hydrophobic organic solvents, water-surfactant, water-ionic liquid, and fluorous/organic solvents. When the mixed homogeneous solutions were delivered into a microspace under certain conditions, the solvent molecules were radially distributed in the microspace, generating inner and outer phases with a kinetic liquid-liquid interface. The TRDP was fundamentally evaluated by fluorescence microscopy, phase diagrams construction, and the elution behaviors of solutes in a capillary tube. A TRDP-based capillary chromatography, referred to as tube radial distribution chromatography (TRDC), where the outer phase serves as a pseudo-stationary phase under laminar flow conditions, has been developed as one of the applications of TRDP. We have also investigated TRDP-based extraction, chemical reaction, and mixing processes, coined as tube radial distribution extraction (TRDE), tube radial distribution reaction (TRDR), and tube radial distribution mixing (TRDM), respectively. The concept and experimental findings regarding TRDP, TRDC, TRDE, TRDR, and TRDM are described in this review.

关键词： Kinetic liquid-liquid interface mixed solvents laminar flow tube radial distribution phenomenon (TRDP) tube radial distribution chromatography (TRDC) tube radial distribution extraction (TRDE) tube radial distribution reaction (TRDR) tube radial distribution mixing (TRDM)

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Phase-separation multiphase flow: preliminary application to analytical chemistry

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ANALYTICAL SCIENCES 2024年第1期40卷 9-28页

作者： tsukagoshi, kazuhiko Doshisha Univ Dept Chem Engn & Mat Sci Kyotanabe Kyoto 6100321 Japan

A two-phase separation mixed solution can undergo phase separation from one phase to two phases (i.e., upper and lower phases) in a batch vessel in response to changes in temperature and/or pressure. This phase separation is reversible. When the mixed solution undergoes a phase change while being fed into a microspace region, a dynamic liquid-liquid interface is formed, leading to a multiphase structure. This flow is called a phase-separation multiphase flow. Annular flow in a microspace, which is one such phase-separation multiphase flow, is interesting and has been applied to chromatography, extraction, reaction fields, and mixing. Here, research papers related to phase-separation multiphase flows-ranging from the discovery of the phenomenon to basic and technical research from the viewpoint of analytical science-are reviewed. In addition, the development of a new separation mode in a high-performance liquid chromatography system based on phase-separation multiphase flow is introduced.

关键词： Phase-separation multiphase flow Tube radial distribution flow (TRDF) Tube radial distribution chromatography (TRDC) Tube radial distribution extraction (TRDE) Tube radial distribution reaction (TRDR) Tube radial distribution mixing (TRDM) Phase-separation mode High-performance liquid chromatography (HPLC)

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Protein separation through preliminary experiments concerning pH and salt concentration by tube radial distribution chromatography based on phase separation multiphase flow using a polytetrafluoroethylene capillary tube

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TALANTA 2017年 169卷 130-135页

作者： Kan, Hyo tsukagoshi, kazuhiko Doshisha Univ Fac Sci & Engn Dept Chem Engn & Mat Sci Kyotanabe Kyoto 6100321 Japan Doshisha Univ Tube Radial Distribut Phenomenon Res Ctr Kyotanabe Kyoto 6100321 Japan

Protein mixtures were separated using tube radial distribution chromatography (TRDC) in a polytetrafluoroethylene (PTFE) capillary (internal diameter=100 mu m) separation tube. Separation by TRDC is based on the annular flow in phase separation multiphase flow and features an open-tube capillary without the use of specific packing agents or application of high voltages. Preliminary experiments were conducted to examine the effects of pH and salt concentration on the phase diagram of the ternary mixed solvent solution of water-acetonitrile-ethyl acetate (8:2:1 volume ratio) and on the TRDC system using the ternary mixed solvent solution. A model protein mixture containing peroxidase, lysozyme, and bovine serum albumin was analyzed via TRDC with the ternary mixed solvent solution at various pH values, i.e., buffer-acetonitrile-ethyl acetate (8:2:1 volume ratio). Protein was separated on the chromatograms by the TRDC system, where the elution order was determined by the relation between the isoelectric points of protein and the pH values of the solvent solution.

关键词： Protein separation Polytetrafluoroethylene capillary tube Phase separation multiphase flow Tube radial distribution flow Tube radial distribution chromatography

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Tube radial distribution chromatography system developed by combining commercially available HPLC system and open-tubular capillary tube as separation column

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TALANTA 2018年 183卷 89-93页

作者： Yamada, Kento Kan, Hyo tsukagoshi, kazuhiko Doshisha Univ Fac Sci & Engn Dept Chem Engn & Mat Sci Kyoto 6100321 Japan Biomicrofluid Sci Res Ctr Kyoto 6100321 Japan

Tube radial distribution chromatography based on tube radial distribution flow, or annular flow, in an open tubular capillary has been reported, where the annular flow is created through phase separation multiphase flow. The chromatographic system requires some specific instruments and treatments for microfluidic flow in the capillary tube. In this study, we developed a new set-up for tube radial distribution chromatography by combining a commercially available HPLC system with an open-tubular capillary tube (with an inner diameter of 100 mu m) as a separation column instead of a conventional packed column. The analyte solution was injected with an injection valve (2 mu L volume) and a ternary solution of water/acetonitrile/ethyl acetate (3:8:2 vol ratio) was delivered as the eluent to the capillary tube at a flow rate of 8.6 mu L min(-1). The chromatographic system, that is, the HPLC system equipped with the open-tubular capillary tube, could successfully separate the model analytes, 1-naphthol, 1-naphthalenesulfonic acid, and 2,6-naphthalenedisulfonic acid, with base-line separation. The inner and outer phases in the annular flow worked as the mobile and pseudo-stationary phases, respectively, in the tube radial distribution chromatography system. The experimentally obtained elution times of the analytes were compared with their corresponding theoretical values calculated using their capacity factors for the inner and outer phases and the linear flow velocities of the respective phases.

关键词： Tube radial distribution chromatography Tube radial distribution flow Phase separation multiphase flow Open-tubular capillary tube HPLC system

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Migration behavior of isoluminol isothiocyanate-labeled α-amino acids in capillary electrophoresis with an absorption/chemiluminescence dual detection system

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JOURNAL OF CHROMATOGRAPHY A 2007年第1-2期1143卷 288-290页

作者： tsukagoshi, kazuhiko Sawanoi, Kaori Nakajima, Rfichiro Doshisha Univ Fac Engn Dept Chem Engn & Mat Sci Kyotanabe 6100321 Japan

In our previous study, we developed capillary electrophoresis with an ultraviolet absorption/chemiluminescence dual detection system. Here, to demonstrate one of the possible applications of the capable system as well as confirm the advantage, migration behavior of isoluminol isothiocyanate-labeled alpha-amino acids was examined in the capillary electrophoresis with a dual detection system. The labeled samples were first analyzed by absorption detection with an on-capillary, followed by chemilunminescence detection with an end-capillary. The system easily, rapidly, and simultaneously produced useful information concerning chemiluminescence quenching and amino group-labeling due to the presence of both absorption and chemiluminescence detections. (c) 2007 Elsevier B.V. All rights reserved.

关键词： capillary electrophoresis chemiluminescence absorption/chemiluminescence dual detection alpha-amino acids

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Development of an immune microanalysis system by use of peroxyoxalate chemiluminescence detection

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ANALYTICAL SCIENCES 2007年第6期23卷 739-741页

作者： tsukagoshi, kazuhiko Tsuge, Kazumasa Nakajima, Riichiro Doshisha Univ Fac Engn Dept Chem Engn & Mat Sci Kyotanabe 6100321 Japan

We developed an immune microanalysis system incorporating chemiluminescence detection, where the peroxyoxalate chemiluminescence (CL) detection using bis[4-nitro-2-(3,6,9-trioxadecyloxycarbonyl)phenyl]oxalate (TDPO)-hydrogen peroxide (H2O2)-fluorescein isothiocianate (FITC) reaction was newly adopted. The analysis system performed the following three processes on a microchip: immune reaction for high selectivity, electrophoresis for formation and transportation of the sample plug, and CL detection. The immune reaction was carried out using an antibody-immobilized glass bead. The glass bead was placed in one of the reservoirs in the microchip along with antigen (analyte) and a known amount of FITC-labeled antigen to set up a competitive immune reaction. The reactant after the immune reaction was fed electrophoretically into the intersection, resulting in a sample plug. The sample plug was then moved into another reservoir containing TDPO-H2O2 acetonitrile solution. At this point, CL detection was performed. The system described here was capable of determining human serum albumin or immunosuppressive acidic protein as a cancer marker in human serum.

关键词： Antigens

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Capillary Chromatography Based on Tube Radial Distribution of Aqueous-Organic Mixture Carrier Solvents: Elution Behavior of Carboxylated Polymer Particles in the System

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JOURNAL OF CHEMICAL ENGINEERING OF JAPAN 2009年第10期42卷 767-770页

作者： Jinno, Naoya Hashimoto, Masahiko tsukagoshi, kazuhiko Doshisha Univ Fac Sci & Engn Dept Chem Engn & Mat Sci Kyotanabe Kyoto 6100321 Japan

A tube radial distribution chromatography (TRDC) system has been developed using an open capillary tube and an aqueous -organic solvent mixture as a carrier solution under laminar flow conditions. In this study, we examine the elution behavior of carboxylated polymer particles (1.0 mu m diameter) together with fluorescein isothiocyanate in the TRDC system in order to extend our knowledge regarding the separation performance of the system. The model analyte solution of fluorescein isothiocyanate and carboxylated polymer particles was injected into the open polyethylene capillary tube by the gravity method, and subsequently delivered through the capillary tube with the carrier solution of the water acetonitrile -ethyl acetate mixture (15:3:2 volume ratio) using a microsyringe pump. On-capillary detection was performed for the analytes using an absorption detector, which indicated individual elution times and peak shapes on the chromatograms. We discuss the observed phenomena considering the separation mechanism of the TRDC system.

关键词： Tube Radial Distribution Chromatography Capillary Tube Laminar Flow Conditions Carboxylate Polymer Particle

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