• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Development of a fully automated lipid analyzer aimed at lipid metabolomics and disease analysis

Research Project

Project/Area Number 16390093
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Pathological medical chemistry
Research InstitutionOsaka University

Principal Investigator

TOJO Hiromasa  Osaka University, Graduate School of Frontier Biosciences, Associate Professor, 生命機能研究科, 助教授 (90135707)

Co-Investigator(Kenkyū-buntansha) TAKEDA Junji  Osaka University, Center for Advanced Science and Innovation, Professor, 先端科学イノベーションセンター, 教授 (50163407)
坂東 泰彦  エーエムアール株式会社, 分析機器部, 部長(研究職)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥15,100,000 (Direct Cost: ¥15,100,000)
Fiscal Year 2005: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 2004: ¥10,100,000 (Direct Cost: ¥10,100,000)
Keywordslipidomics / sphingolipids / phospholipids / HPLC / mass spectrometry / リピドミックス / 脂質 / 順相クロマトグラフィー / エレクトロスプレー
Research Abstract

Lipidomics has attracted much attention because a side-by-side analysis of proteome and lipid metabolome could greatly facilitate to understand protein functions relevant to lipid metabolism. We developed an automated normal-phase microbore HPLC/electrospray (ESI) ion trap mass spectrometry (MS) system for analyzing a wider range of lipid classes, from neutral lipids (e.g., cholesterol esters, waxes, glycerides, ceramides, and glycosphingolipids) to phospholipids, on a single chromatographic run. The system uses a ternary gradient and post-column addition of solvent that aids ESI of neutral lipids eluted earlier with non-polar solvents. A pulse-less pump that can stably deliver gradients of non-polar solvents at low flow rates was developed. To increase ionization efficiency, the recently developed fluoropolymer-coated ESI tip, FortisTip, was used, and the effluent was dispersed via a T splitter, yielding a flow rate of 〜0.5μl/min across the tip, and monitored with a Finnigan LCQdeca-X … More P spectrometer set to either data dependent negative or positive tandem MS^2 and MS^3 mode as well as to alternate positive and negative ion full scan (alternate polarity switching) mode. The other effluent at 〜50μl/min was monitored with an evaporative light scattering photometer, which is useful for detecting cholesterol on-line. Choresterol ester, triacylglycerol, ceramide, cerebroside, phosphatidylethanolamine, phosphatidylcholine, and sphingomyelin with acyl chain(s) of unnatural length were used as internal standards. The MS-based chromatographic profiling, encompassing neutral lipids and phospholipids, is useful for detecting and structurally identifying a novel lipid and lipid anomaly in diseased states. This method was successfully applied in response to real analytical needs, including determination of changes in lipid profile in mice with tissue-specific disruption of sphingolipid metabolism-related gene, such as serine palmitoyl transferase (SPT), yeast lipid profile changes on gene manipulation, bacterial SPT activity assay, analysis of a novel glucose-containing phospholipid, phosphatidylglucoside, in brain, and lipid analysis in the active site of acyl CoA oxidase in single crystals. Less

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (22 results)

All 2006 2005 2004 Other

All Journal Article (22 results)

  • [Journal Article] Three-Dimensional Structure of Rat-Liver Acyl-CoA Oxidase in Complex with Fatty Acid : Insights into Substrate-ecognition and Reactivity toward Molecular Oxygen2006

    • Author(s)
      Tokuoka, K.
    • Journal Title

      J.Biochem. 139

      Pages: 789-795

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Analysis and determination of novel glycolipids in vertebrate brains by HPLC/mass spectreometry.2006

    • Author(s)
      Nagatsuka, Y., Tojo, H., Hirabayashi, Y.
    • Journal Title

      Methods Enzymol (In press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Three-Dimensional Structure of Rat-Liver Acyl-CoA Oxidase in Complex with a Fatty Acid : Insights into Substrate-Recognition and Reactivity toward Molecular Oxygen2006

    • Author(s)
      Tokuoka, K., Nakajima, Y., Hirotsu, K., Miyahara, I., Nishina Y., Shiga, K., Tamaoki, H., Setoyama, C., Tojo, H., Miura, R.
    • Journal Title

      J.Biochem. 139

      Pages: 789-795

    • NAID

      10018845985

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A cytosolic cytochrome b5-like protein in yeast cell accelerating the electron transfer from NADPH to cytochrome c catalyzed by Old Yellow Enzyme.2005

    • Author(s)
      Nakagawa, M.
    • Journal Title

      Biochem.Biophys.Res.Commun. 338

      Pages: 605-609

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] The Angiotensin-converting Enzyme (ACE) is a GPI-anchored Protein Releasing Factor Crucial for Fertilization.2005

    • Author(s)
      Kondo, G.
    • Journal Title

      Nature Med. 11

      Pages: 160-166

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] リパーゼファミリーに属するホスホリパーゼA2 触媒トライアッドの不思議2005

    • Author(s)
      東城博雅
    • Journal Title

      化学と生物 43

      Pages: 516-518

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A cytosolic cytochrome b5-like protein in yeast cell accelerating the electron transfer from NADPH to cytochrome c catalyzed by Old Yellow Enzyme.2005

    • Author(s)
      Nakagawa, M., Yamano, T., Kuroda, K., Nonaka, Y., Tojo, H., Fujii, S.
    • Journal Title

      Biochem.Biophys.Res.Commun. 338

      Pages: 605-609

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] The Angiotensin-converting Enzyme (ACE) is a GPI-anchored Protein Releasing Factor Crucial for Fertilization.2005

    • Author(s)
      Kondo, G., Tojo, H., Nakatani, Y., Komazawa, N., Murata, C., Yamagata, K., Maeda, Y., Kinoshita, T., Okabe, M., Taguchi, R., Takeda, J.
    • Journal Title

      Nature Med. 11

      Pages: 160-166

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Phospholipase A2 belonging to lipase families.2005

    • Author(s)
      Tojo, H.
    • Journal Title

      KASEAA 43

      Pages: 516-518

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] リパーゼファミリーに属するホスホリパーゼA2 触媒トライアッドの不思議2005

    • Author(s)
      東城 博雅
    • Journal Title

      化学と生物 43

      Pages: 516-518

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Fully automated online multi-dimensional protein profiling system for complex mixtures.2004

    • Author(s)
      Fujii, K.
    • Journal Title

      J.Chromatogr.A 1057

      Pages: 107-113

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Properties of an electrospray emitter coated with material of low surface energy.2004

    • Author(s)
      Tojo, H.
    • Journal Title

      J.Chromatogr.A 1056

      Pages: 223-228

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 質量分析を利用した脂質メタボロミックスの現況2004

    • Author(s)
      東城博雅
    • Journal Title

      オレオサイエンス 4

      Pages: 3-12

    • NAID

      10012704729

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary 2004 Annual Research Report
  • [Journal Article] Recent developments in mass spectrometry-based lipid metabolomics2004

    • Author(s)
      Tojo, H.
    • Journal Title

      Oleoscience 4

      Pages: 147-156

    • NAID

      10012704729

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Properties of an electrospray emitter coated with material of low surface energy.2004

    • Author(s)
      Tojo, H.
    • Journal Title

      J.Chromatogr. A1056

      Pages: 223-228

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Fully automated online multi-dimensional protein profiling system complex mixtures2004

    • Author(s)
      Fujii, K., Nakano, T., Hike, H., Usui, F., Bando, Y., Tojo, H., Nishimura, T.
    • Journal Title

      J.Chromatogr. A1057

      Pages: 107-113

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] The Angiotensin-converting Enzyme (ACE) is a GPI-anchored Protein Releasing Factor Crucial for Fertilization2004

    • Author(s)
      Kondoh, G et al.
    • Journal Title

      Nature Med. (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Properties of an electrospray emitter coated with material of low surface energy2004

    • Author(s)
      Tojo, H
    • Journal Title

      J.Chromatogr.A 1056

      Pages: 223-228

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Fully automated online multi-dimensional protein profiling system for complex mixtures2004

    • Author(s)
      Fuji, K.et al.
    • Journal Title

      J.Chromatogr.A 1057

      Pages: 107-113

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Analysis and determination of novel glycolipids in vertebrate brains by HPLC/mass spectrometry.

    • Author(s)
      Nagatsuka, Y.
    • Journal Title

      Methods Enzymol. (印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Analysis and determination of novel glycolipids in vertebrate brains by HPLC/mass spectrometry.

    • Author(s)
      Nagatsuka, Y.
    • Journal Title

      Methods Enzymol. (印刷中)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Three-Dimensional Structure of Rat-Liver Acyl-CoA Oxidase in Complex with Fatty Acid : Insights into Substrate-Recognition and Reactivity toward Molecular Oxygen

    • Author(s)
      Tokuoka, K.
    • Journal Title

      J.Biochem. (印刷中)

    • Related Report
      2005 Annual Research Report

URL: 

Published: 2004-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi