Chemical Speciation of Arsenic Species in Human Blood Serum by Liquid Chromatography Using a Phosphatidylcholine-Coated ODS Column with Detection by ICP-MS

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  • Hasegawa Takuya
    Department of Applied Chemistry, Graduate School of Engineering, Nagoya University
  • Ishise Jotaro
    Department of Applied Chemistry, Graduate School of Engineering, Nagoya University
  • Fukumoto Yasuharu
    Department of Applied Chemistry, Graduate School of Engineering, Nagoya University
  • Matsuura Hirotaka
    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
  • Yamamoto Kazuhito
    Department of Hematology and Oncology, Graduate School of Medicine, Nagoya University
  • Naoe Tomoki
    Department of Hematology and Oncology, Graduate School of Medicine, Nagoya University

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Abstract

Chemical speciation of arsenic species in human blood serum was performed by high-performance liquid chromatography/inductively coupled plasma mass spectrometry (HPLC/ICP-MS) with direct sample injection, where an octadecylsilyl silica (ODS) column coated with phosphatidylcholine (PC) (hereafter known as “PC-coated ODS column”) was used as the separation column. In arsenic species analysis, a citrate buffer solution (pH 4.0) was used as the mobile phase, in which the following reagents were added: sodium 1-dodecanesulfonate (SDS), tetramethylammonium hydroxide (TMAH), which are ion-pair reagents to separate inorganic and organic arsenic species, and 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS), which is a protein-solubilizing agent to prevent adsorption of proteins on the column. As a result of optimization, five representative arsenic species spiked in human blood serum reference material could be separated from each other within 5 min on the PC-coated ODS column by elution with a 5 mM citrate buffer (pH 4.0) containing 5 mM SDS, 5 mM THAH, and 0.2 mM CHAPS. The detection limits obtained by ICP-MS were 3.1, 2.7, 4.5, 2.5, and 2.5 ng of As g−1 for arsenate (iAsV), arsenite (iAsIII), monomethylarsonic acid (MMA), dimethylarsenic acid (DMA), and arsenobetaine (AB), respectively, when the injection volume was 20 μL. The present separation system was also applied to speciation analysis of arsenic species in human blood serum collected from a leukemia patient after therapeutic treatment with arsenic.

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