分子インプリント撹拌子による生体内環境汚染物質の高感度分析と暴露評価に関する研究

• Project/Area Number: 04J01353
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 国内
• Research Field: Analytical chemistry
• Research Institution: Hoshi University
• Principal Investigator: 川口 研 星薬科大学, 大学院薬学研究科, 特別研究員DC1
• Project Period (FY): 2004 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥2,800,000 (Direct Cost: ¥2,800,000); Fiscal Year 2006: ¥900,000 (Direct Cost: ¥900,000); Fiscal Year 2005: ¥900,000 (Direct Cost: ¥900,000); Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
• Keywords: 分子インプリント法 / 安定同位体 / MIP / IMIP / MISBE / SBSE / マイクロ抽出 / BPA

Research Abstract

わずか一滴の生体試料から様々な情報を得ることが可能になれば、化学物質の暴露と健康影響の関係を調査や病気の診断等を容易に行うことができる。とりわけ、化学物質の影響を受けやすい胎児や新生児の生体試料は稀少であり、超高感度分析法の開発が必要とされている。本研究の目的は、分子インプリント(MI)法を用いた生内微量化学物質の超高感度分析法の開発にある。MI法は、認識させたい分子(鋳型分子)とモノマーを重合し、得られたポリマーから鋳型分子を取り除くことにより、鋳型分子に対して相補的な結合部位を形成する方法である。得られたポリマー(MIP)は、鋳型分子を選択的に吸着することが可能となる。
一昨年度の研究において、従来のMI法の欠点であった作製したポリマー内部からの分析対象物質の溶出を克服した新しいMIPとして、分析対象物質の安定同位体ラベル化物質を鋳型分子とするIMIPの作製に成功した。
また、昨年度の研究では、作製したIMIPとガラス製の撹搾子を限外ろ過膜で包んだ分子インプリント撹搾子(MISB)を開発した。
本年度は、作製したMISBを用いて、実際にヒト生体試料である尿及び血漿中ビスフェノールA(BPA)の前処理を行った。脱抱合体化処理を行った尿の分析において、sub ng/mlレベルでの高感度な測定を可能とした。しかし、血漿の測定では、マトリックス(アルブミン等)の影響を受けて、満足できる回収率を得ることは出来なかった。今後は、IMIPの合成において、ポリマー表面を親水化した浸透制限型分子インプリントポリマーを作製することで、アルブミン等のタンパクの吸着を防ぐ手法が有効であると示唆された。

Research Products

• [Journal Article] Liquid phase microextraction with in situ derivatization for measurement of bisphenol A in river water sample by gas chromatography-mass spectrometry (2006)
  • Author(s): M.Kawaguchi et al.
  • Journal Title: J. Chromatogr. A 1110 Pages: 1-5
• [Journal Article] Novel stir bar sorptive extraction methods for environmental and biomedical analysis (2006)
  • Author(s): M.Kawaguchi et al.
  • Journal Title: J. Pharm. Biomed. Anal. 40 Pages: 500-508
• [Journal Article] Stir bar sorptive extraction and thermal desorption-gas chromatography-mass spectrometry for trace analysis of benzophenone and its derivatives in water sample (2006)
  • Author(s): M.Kawaguchi et al.
  • Journal Title: Anal. Chim. Acta 557 Pages: 272-277
• [Journal Article] Dual derivatization-stir bar sorptive extraction-thermal desorption-gas chromatography-mass spectrometry for the determination of l7 β-Estradiol in water sample (2006)
  • Author(s): M.Kawaguchi et al.
  • Journal Title: J. Chromatogr. A 1105 Pages: 140-147
• [Journal Article] Stir bar sorptive extraction with in situ de-conjugation and thermal desorption gas chromatography-mass spectrometry for measurement of 4-nonylphenol glucuronide in human urine sample (2006)
  • Author(s): M.Kawaguchi et al.
  • Journal Title: J. Pharm. Biomed. Anal. 40 Pages: 82-87
• [Journal Article] Dual derivatization-stir bar sorptive extraction-thermal desorption-gas chromatography-mass spectrometry for determination of 17β-Estradiol in water sample (2006)
  • Author(s): M.Kawaguchi et al.
  • Journal Title: J.Chromatogr.A 1105 Pages: 140-147
• [Journal Article] Stir bar sorptive extraction and thermal desorption-gas chromatography-mass spectrometry for trace analysis of benzophenone and its derivatives in water sample (2006)
  • Author(s): M.Kawaguchi et al.
  • Journal Title: Anal.Chim.Acta 557 Pages: 272-277
• [Journal Article] Molecularly imprinted solid phase extraction using stable isotope labeled compounds as template and liquid chromatography-mass spectrometry for trace analysis of bisphenol A in water sample (2005)
  • Author(s): M.Kawaguchi et al.
  • Journal Title: Anal.Chim.Acta 539 Pages: 83-89
• [Journal Article] Stir bar sorptive extraction with in situ derivatization and thermal desorption-gas chromatography-mass spectrometry for measurement of phenolic xenoestrogens in human urine samples (2005)
  • Author(s): M.Kawaguchi et al.
  • Journal Title: J.Chromatogr.B 820 Pages: 49-57
• [Journal Article] Stir bar sorptive extraction with in situ de-conjugation and thermal desorption gas chromatography-mass spectrometry for measurement of 4-nonylphenol glucuronide in human urine sample (2005)
  • Author(s): M.Kawaguchi et al.
  • Journal Title: J.Pharm.Biomed.Anal 60 Pages: 82-87
• [Journal Article] Stir bar sorptive extraction and trace analysis of alkylphenols in water samples by thermal desorption with in tube silylation and gas chromatography-mass spectrometry (2005)
  • Author(s): Migaku Kawaguchi
  • Journal Title: J.Chromatogr.A 1062 Pages: 23-29
• [Journal Article] Stir bar sorptive extraction with in situ derivatization and thermal desorption-gas chromatography-mass spectrometry for determination of chlorophenols in water and body fluid samples (2005)
  • Author(s): Migaku Kawaguchi
  • Journal Title: Anal.Chim.Acta 533 Pages: 57-65
• [Journal Article] Determination of bisphenol A in river water and body fluid samples by stir bar sorptive extraction with in situ derivatization and thermal desorption-gas chromatography-mass spectrometry (2004)
  • Author(s): Migaku Kawaguchi
  • Journal Title: J.Chromatogr.B 805 Pages: 41-48
• [Journal Article] Trace analysis of phenolic xenoestrogens in water samples by stir bar sorptive extraction with in situ derivatization and thermal desorption-gas chromatography-mass spectrometry (2004)
  • Author(s): Migaku Kawaguchi
  • Journal Title: J.Chromatogr.A 1041 Pages: 19-26
• [Journal Article] Determination of 4-tert-octylphenol and 4-nonylphenol in laboratory animal feed sample by stir bar sorptive extraction followed by liquid desorption and column switching-liquid chromatography-mass spectrometry with solid phase extraction (2004)
  • Author(s): Migaku Kawaguchi
  • Journal Title: J.Chromatogr.A 1046 Pages: 83-88
• [Journal Article] Trace analysis of chlorophenols in river water samples by stir bar sorptive extraction with in situ derivatization and thermal desorption-gas chromatography-mass spectrometly (2004)
  • Author(s): Migaku Kawaguchi
  • Journal Title: Organohalogen Compounds 66 Pages: 181-185
• [Journal Article] Stir bar sorptive extraction with in situ derivatization and thermal desorption-gas chromatography-mass spectrometry in the multi shot mode for determination of estrogens in river water samples (2004)
  • Author(s): Migaku Kawaguchi
  • Journal Title: J.Chromatogr.A 1049 Pages: 1-8
• [Journal Article] Stir-bar-sorptive extraction, with in-situ deconjugation, and thermal desorption with in-tube silylation, followed by gas chromatography-mass spectrometry for measurement of urinary 4-nonylphenol and 4-tert-octylphenol glucuronides
  • Author(s): M.Kawaguchi et al.
  • Journal Title: Anal. Bioanal. Chem. (in press)
• [Journal Article] Novel stir bar sorptive extraction methods for environmental and biomedical analysis
  • Author(s): M.Kawaguchi et al.
  • Journal Title: J.Pharm.Biomed.Anal. (in press)
• [Journal Article] Molecularly imprinted solid phase extraction using stable isotope labeled compounds as template and liquid chromatography-mass spectrometry for trace analysis of bisphenol A in water sample
  • Author(s): Migaku Kawaguchi
  • Journal Title: Anal.Chim.Acta (in press)
• [Journal Article] Stir bar sorptive extraction with in situ derivatization and thermal desorption-gas chromatography-mass spectrometry for measurement of phenolic xenoestrogens in human urine samples
  • Author(s): Migaku Kawaguchi
  • Journal Title: J.Chromatogr.B (in press)