Biomolecule Analyses in an Open-Tubular Capillary Chromatography Using Ternary Mixed Carrier Solvents with Chemiluminescence Detection

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  • TAKAHASHI Naoya
    Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University
  • MASUHARA Yuji
    Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University
  • JINNO Naoya
    Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University
  • HASHIMOTO Masahiko
    Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University
  • TSUKAGOSHI Kazuhiko
    Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University

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We examined the elution behavior of isoluminol isothiocyanate (ILITC)-labeled biomolecules (α-amino acids, peptides, and proteins) in an open-tubular capillary chromatography system using an untreated fused-silica capillary tube and a water–acetonitrile–ethyl acetate mixture carrier solution. Such an open-tubular capillary chromatography is called “tube radial distribution chromatography (TRDC)” for convenience. A mixture of ILITC and ILITC-labeled biomolecules was analyzed using TRDC with chemiluminescence detection that provided simple instrument without a light source and complex optical devises. The ILITC and the labeled twenty α-amino acids were separated, in this order or the reverse order, or not separated with an organic solvent-rich and water-rich carrier solution. Their elution behavior was considered to be of hydrophilic or hydrophobic nature of ILITC and the labeled α-amino acids. The ILITC and the labeled protein, alcohol dehydrogenase and bovine serum albumin, were separated in this order with an organic solvent-rich carrier solution, while they were eluted in the reverse order with a water-rich carrier solution, based on the TRDC separation performance. The TRDC system worked with the untreated open-tubular capillary tube not using any specific capillary tubes, such as coated, packed, or monolithic.

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  • Analytical Sciences

    Analytical Sciences 28 (4), 351-351, 2012

    社団法人 日本分析化学会

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