Separation Characteristics of a Phosphatidylcholine-Coated ODS Column for Direct Sample Injection Analysis of Biological Fluid Samples

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  • Hasegawa Takuya
    Department of Applied Chemistry, Graduate School of Engineering, Nagoya University
  • Fukumoto Yasuharu
    Department of Applied Chemistry, Graduate School of Engineering, Nagoya University
  • Ishise Jotaro
    Department of Applied Chemistry, Graduate School of Engineering, Nagoya University
  • Hattori Ryota
    Department of Applied Chemistry, Graduate School of Engineering, Nagoya University
  • Zhu Yanbei
    Department of Applied Chemistry, Graduate School of Engineering, Nagoya University
  • Umemura Tomonari
    Department of Applied Chemistry, Graduate School of Engineering, Nagoya University
  • Haraguchi Hiroki
    Department of Applied Chemistry, Graduate School of Engineering, Nagoya University

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Abstract

An octadecylsilyl silica (ODS) column coated with phosphatidylcholine (PC) was prepared by using a dynamic coating method, and its separation characteristics were evaluated for direct sample injection analysis of biological fluids. First, fundamental separation behaviors of various kinds of large and small molecules/ions were examined by HPLC using the PC-coated ODS column with UV absorption detection, for which pure water was used as a mobile phase. In the experiments, proteins (large molecules) were rapidly eluted in the range of 2.07 min (dead time)–3.20 min, while small molecules, such as inorganic anions, amino acids, organic acids, and organic drugs, were eluted after 3.50 min. The fact that proteins and small molecules/ions were separated on the PC-coated ODS column suggested that the present column can be used for direct sample injection analysis of biological fluid samples, such as urine and blood serum. Thus, the PC-coated ODS column was used in the analysis of arsenic species in urine and in the analysis of nicotinamide, theophylline, tryptophan, and caffeine in a model blood serum solution without any sample pretreatment.

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