Screening of inherited diseases with abnormal sterol metabolism by LC-MS/MS

2010 Fiscal Year Final Research Report

• Project/Area Number: 20591309
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Embryonic/Neonatal medicine
• Research Institution: Tokyo Medical University
• Principal Investigator: HONDA Akira Tokyo Medical University, 医学部, 准教授 (10468639)
• Co-Investigator(Kenkyū-buntansha): MATSUZAKI Yasushi 東京医科大学, 医学部, 教授 (50209532) ; IKEGAMI Tadashi 東京医科大学, 医学部, 准教授 (40439740)
• Project Period (FY): 2008 – 2010
• Keywords: 先天異常学 / ステロール代謝 / 生化学的診断 / 血清診断 / 高速液体クロマトグラフィー / タンデムマススペクトロメトリー

Research Abstract

We have developed a highly sensitive and specific method for the analysis of sterol profiles in biological samples. Sterols in less than 10μl of dried serum were derivatized into picolinyl esters and analyzed by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) using the electrospray ionization (ESI) mode. In addition to cholesterol, more than 50 cholesterol related sterols were simultaneously identified within 50 minutes. Quantitative analyses for the picolinyl esters of 20 available sterols were performed. Detection limits were found to be less than 1 pg on-column, and sufficient reproducibilities and recoveries of these sterols were observed. The present method provides reliable and reproducible results for the identification and quantification of neutral sterols, especially in small volumes of blood samples, which is useful for serological diagnosis of inherited diseases with abnormal sterol metabolism.

Research Products

• [Journal Article] Cholesterol 25-hydroxylation activity of CYP3A (2011)
  • Author(s): Honda A, Miyazaki T, Ikegami T, Iwamoto J, Maeda T, Hirayama T, Saito Y., Teramoto T, Matsuzaki Y
  • Journal Title: J. Lipid Res(Epub ahead of print)
  • Peer Reviewed
• [Journal Article] Cholesterol and chronic hepatitis C infection (2011)
  • Author(s): Honda A, Matsuzaki Y
  • Journal Title: Hepatol. Res(In press)
  • Peer Reviewed
• [Journal Article] Highly sensitive and specific analysis of sterol profiles in biological samples by HPLC-ESI-MS/MS. (2010)
  • Author(s): Honda A, Miyazaki T, Ikegami T, Iwamoto J, Yamashita K, Numazawa M, Matsuzaki Y
  • Journal Title: J. Steroid Biochem. Mol. Biol 121 Pages: 556-564
  • Peer Reviewed
• [Journal Article] A SNP of NPC1L1 affects cholesterol absorption in Japanese. (2010)
  • Author(s): Maeda T, Honda A, Ishikawa T, Kinoshita M, Mashimo Y, Takeoka Y, Yasuda D, Kusano J, Tsukamoto K, Matsuzaki Y, Teramoto T
  • Journal Title: J. Atheroscler. Thromb 17 Pages: 356-360
  • Peer Reviewed
• [Journal Article] Highly sensitive quantification of key regulatory oxysterols in biological samples by LC-ESI-MS/MS (2009)
  • Author(s): Honda A, Yamashita K, Hara T, Ikegami T, Miyazaki T, Shirai M, Xu G, Numazawa M, Matsuzaki Y
  • Journal Title: J. Lipid Res 50 Pages: 350-357
  • Peer Reviewed
• [Journal Article] Highly-sensitive quantification of serum malonate, a possible marker for de novo lipogenesis, by LC-ESI-MS/MS (2009)
  • Author(s): Honda A, Yamashita K, Ikegami T, Hara T, Miyazaki T, Hirayama T, Numazawa M, Matsuzaki Y
  • Journal Title: J. Lipid Res. 50 Pages: 2124-2130
  • Peer Reviewed
• [Journal Article] Highly sensitive analysis of sterol profiles in human serum by LC-ESI-MS/MS. (2008)
  • Author(s): Honda A, Yamashita K, Miyazaki H, Shirai M, Ikegami T, Xu G, Numazawa M, Hara T, Matsuzaki Y
  • Journal Title: J. Lipid Res 49 Pages: 2063-2073
  • Peer Reviewed
• [Journal Article] Determination of key intermediates in cholesterol and bile acid biosynthesis by stable isotope dilution mass spectrometry. (2008)
  • Author(s): Yoshida T, Honda A, Miyazaki H, Matsuzaki Y
  • Journal Title: Anal. Chem. Insights 3 Pages: 45-60
  • Peer Reviewed
• [Presentation] 原発性胆汁性肝硬変に対するUDCAおよびベザフィブラートによる治療効果発現メカニズムの比較 (2010)
  • Author(s): 本多彰, 池上正, 中牟田誠, 宮崎照雄, 齋藤吉史, 平山剛, 松崎靖司
  • Organizer: 第46回日本肝臓学会総会(ワークショップ)
  • Place of Presentation: 山形
  • Year and Date: 2010-05-27
• [Presentation] HPLC-MS/MSを用いた先天性ステロール代謝異常症一斉診断システムの確立 (2009)
  • Author(s): 本多彰, 宮崎照雄, 池上正, 原孝, 松崎靖司
  • Organizer: 第51回日本先天代謝異常学会総会
  • Place of Presentation: 東京
  • Year and Date: 2009-11-07
• [Presentation] Non-invasive measurement of cholesterol metabolism in patients with HCV infection by using serum marker sterols (2009)
  • Author(s): Ikegami T, Honda A, Nakamuta M, Miyazaki T, Hirayama T, Saitoh Y, Matsuzaki Y.
  • Organizer: 第60回米国肝臓病学会
  • Place of Presentation: Boston
  • Year and Date: 2009-11-02
• [Presentation] 原発性胆汁性肝硬変におけるコレステロール代謝の解析 (2009)
  • Author(s): 本多彰,中牟田誠,松崎靖司
  • Organizer: 第51回日本消化器病学会大会
  • Place of Presentation: 京都
  • Year and Date: 2009-10-16
• [Presentation] コレステロール・胆汁酸代謝系に関するメタボローム解析方法の開発 (2008)
  • Author(s): 本多彰, 池上正, 宮崎照雄, 松崎靖司
  • Organizer: 第12回日本肝臓学会大会
  • Place of Presentation: 東京
  • Year and Date: 20081001-20081003
• [Book] Encyclopedia of Molecular Mechanisms of Disease(F. Lang ed.) (Cerebrotendinous Xanthomatosis) (2009)
  • Author(s): Honda A
  • Total Pages: 310-311
  • Publisher: Springer
• [Remarks] ホームページ等