Ultrafast determination of drugs in biological samples by column-switching LC/MS

• Project/Area Number: 19590683
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Legal medicine
• Research Institution: Aichi Medical University
• Principal Investigator: ARINOBU Tetsuya Aichi Medical University, 医学部, 准教授 (30329776)
• Project Period (FY): 2007 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2009: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2008: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 法中毒学 / 迅速分析 / 分析科学 / 有機化学 / 質量分析 / 高速液体クロマトグラフィー / 血中薬物

Research Abstract

Theophylline and caffeine are natural alkaloids widely being ingested, and show various pharmacological effects in humans. Caffeine is a constituent of coffee and other beverages and included in many medicines. Theophylline is used as a bronchodilator and also formed as a metabolite of caffeine in humans ; theophylline poisoning is occasionally encountered in the field of forensic toxicology, because of its relatively narrow safe range of therapeutic doses. They have been analyzed by various methods, such as gas chromatography (GC), GC-mass spectrometry (MS), high-performance (HP) liquid chromatography (LC)-MS. In the existing analytical methods for the drugs, the sample pretreatments such as liquid-liquid extraction, solid-phase extraction, filtration or centrifugation were usually required to remove proteins and other impurities contained in biological matrices. The analytical methods including time-consuming multi-step sample pretreatments are unsuitable for rapid detection of poisonous materials in human specimens. To avoid such sample pretreatments, special HPLC columns, which enabled direct injection of crude biological samples, were developed ; our group applied them to compounds of forensic toxicological interest. Furthermore, backflush column-switching system using one of special HPLC columns have become a powerful analytical tool for simultaneous determinations of various drugs in biological samples. In this study, we have established an automated on-line method for ultra-fast determination of theophylline and caffeine in human serum by HPLC-MS with backflush column-switching and a monolithic separation column. To our knowledge, our method presented in this study is the fastest MS determination technique for theophylline and caffeine in human serum using the conventional HPLC instrument.

Research Products

• [Journal Article] High-throughput determination of theophylline and caffeine in human serum by conventional liquid chromatography-mass spectrometry. (2009)
  • Author(s): Arinobu T, Hattori H, Kumazawa T, Lee X-P, Mizutani Y, Katase T, Kojima S, Omori T, Kaneko R, Ishii A, Seno H.
  • Journal Title: Forensic Toxicol. 27 Pages: 1-6
  • Peer Reviewed
• [Journal Article] High-throughput determination of theophylline and caffeine in human serum by conventional liquid chromatography-mass spectrometry (2009)
  • Author(s): Tetsuya Arinobu, Hideki Hattori, Takeshi Kumazawa, Xiao-Pen Lee, Yoko Mizutani, Takao Katase, Sadao Kojima, Takayuki Omori, Rina Kaneko, Akira Ishii, Hiroshi Seno
  • Journal Title: Forensic Toxicology 27 Pages: 1-6
  • Peer Reviewed
• [Journal Article] An automated on-line method for simultaneous analysis of phenothiazines in human serum by high-performance liquid chromatography/sonic spray ionization mass spectrometry using backflush column switching. (2007)
  • Author(s): Aoki R, Arinobu T, Kumazawa T, Hattori H, Noguchi H.
  • Journal Title: Forensic Toxicol. 25(1) Pages: 8-15
  • Peer Reviewed
• [Journal Article] Rapid analysis of sertraline, fluvoxamine, and paroxetine in serum specimens by LC-MS-MS using a new polymer column. (2007)
  • Author(s): Hattori H, Ito K, Iwai M, Arinobu T, Mizutani Y, Kumazawa T, Ishii A, Suzuki O, Seno H.
  • Journal Title: Forensic Toxicol. 25(2) Pages: 100-103
  • Peer Reviewed
• [Journal Article] An automated on-line method for simultaneous analysis of phenothiazines in human serum by high-performance liquid chromatography/sonic spray ionization mass spectrometry using backflush column switching (2007)
  • Author(s): Ruri Aoki, Tetsuya Arinobu, Takeshi Kumazawa, Hideki Hattori, Hiroshi Noguchi
  • Journal Title: Forensic Toxicology 25 Pages: 8-15
  • Peer Reviewed
• [Journal Article] Rapid analysis of sertraline, fluvoxamine, and paroxetine in serum specimens by LC-MS-MS using a new polymer column (2007)
  • Author(s): Hideki Hattori, Kenjiro Ito, Masae Iwai, Tetsuya Arinobu, Yoko Mizutani, Takeshi Kumazawa, Akira Ishii, Osamu Suzuki, Hiroshi Seno
  • Journal Title: Forensic Toxicology 25 Pages: 100-103
  • Peer Reviewed
• [Presentation] モノリスカラムを用いたバックフラッシュカラムスイッチングLC/MSによる血中テオフィリンの超迅速分析 (2009)
  • Author(s): 有信哲哉, 服部秀樹, 熊澤武志, 李暁鵬, 水谷洋子, 大森貴之, 西本寛, 小島貞男, 石井晃, 妹尾洋
  • Organizer: 第34回日本医用マススペクトル学会
  • Place of Presentation: 近畿大学(大阪府)
  • Year and Date: 2009-09-10
• [Presentation] モノリスカラムを用いたバックフラッシュカラムスイッチングLC/MSによる血中テオフィリンの超迅速分析 (2009)
  • Author(s): 有信哲哉, 服部秀樹, 熊澤武志, 李暁鵬, 水谷洋子, 大森貴之, 西本寛, 小島貞男, 石井晃, 妹尾洋
  • Organizer: 第34回日本医用マススペクトル学会年会
• [Presentation] イオンペア試薬を用いた尿中第四級アンモニウム塩系筋弛緩剤の分析 (2008)
  • Author(s): 伊藤健次郎, 服部秀樹, 有信哲哉, 妹尾洋
  • Organizer: 第90次日本法医学会総会
• [Presentation] 血清中SNRI, DNRIおよびNRIのLS/MS/MSによる迅速分析 (2008)
  • Author(s): 服部秀樹, 伊藤健次郎, 金子理奈, 有信哲哉, 岩井雅枝, 水谷洋子, 石井晃, 鈴木修, 妹尾洋
  • Organizer: 日本法中毒学会
  • Place of Presentation: 昭和大学
• [Presentation] Determination of phenothiazines in human serum by HPLC/SSI-MS using backflush column-switching. (2007)
  • Author(s): Arinobu T, Aoki R, Kumazawa T, Lee X-P., Ishii A, Kaneko R, Kojima S, Noguchi H, Suzuki O, Hattori H, Seno H
  • Organizer: 12^<th> International meeting on recent developments in pharmaceutical analysis
  • Place of Presentation: Elba, Italy
• [Presentation] DETERMINATION OF PHENOTHIAZINES IN HUMAN SERUM BY HPLC/SSI-MSUSING BACKFLUSH COLUMN-SWITCHING (2007)
  • Author(s): T. Arinobu, R. Aoki, T. Kumazawa, X-P. Lee, A. Ishii, R. Kaneko, S. Kojima, H. Noguchi, O. Suzuki, H. Hattori, H. Seno
  • Organizer: 12th International meeting on recent developments in pharmaceutical analysis
  • Place of Presentation: Elba, Italy