Development of a lipid profiling system using reverse-phase liquid chromatography coupled to high-resolution mass spectrometry with rapid polarity switching and an automated lipid identification software

Takayuki Yamada, Takato Uchikata, Shigeru Sakamoto, Yasuto Yokoi, Eiichiro Fukusaki, Takeshi Bamba

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

Lipidomics requires accurate lipid profiling, which until recently has been challenging at best. In the present study, we developed a practical workflow for high-throughput and exhaustive lipid profiling by combining reverse-phase liquid chromatography coupled to quadrupole orbitrap Fourier transform mass spectrometry, with an automated lipid identification software. This validated method enables highly sensitive and simultaneous analysis of lipids with varying polarities such as glycerophospholipids and sphingophospholipids, by switching the acquisition polarities in mass spectrometry. In addition, it facilitates data-dependent MS2 analysis targeting the lipid molecular species without any influence from other ions by setting the inclusion list, the target m/z list for the product ion scanning. The m/z values of the target lipid molecular species, stored in the database of Lipid Search software, are added to the inclusion list. Moreover, optimizing the identification conditions of the software for the LC/MS system enables high-throughput and accurate identification of lipid molecular species existing in biological samples. Specifically, LC separation is essential for accurate identification of lipid molecular species that possess some fatty acid chains, because it can be difficult to determine fatty acid chain composition of detected molecular species especially in triacylglycerol compounds in direct infusion mass spectrometry. This method has high reproducibility and can be used for structural analysis even for low-abundance compounds. Using this method, over 400 lipid compounds targeted in this research were detected and identified from a sample of mouse plasma. This result indicates that the LC/MS method in the present study enables efficient lipid profiling.

Original languageEnglish
Pages (from-to)211-218
Number of pages8
JournalJournal of Chromatography A
Volume1292
DOIs
Publication statusPublished - May 31 2013
Externally publishedYes

Fingerprint

Liquid chromatography
Reverse-Phase Chromatography
Mass spectrometry
Mass Spectrometry
Software
Lipids
Fatty Acids
Throughput
Ions
Glycerophospholipids
Workflow
Fourier Analysis
Structural analysis
Fourier transforms
Triglycerides
Databases
Scanning
Plasmas

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Organic Chemistry
  • Biochemistry

Cite this

Development of a lipid profiling system using reverse-phase liquid chromatography coupled to high-resolution mass spectrometry with rapid polarity switching and an automated lipid identification software. / Yamada, Takayuki; Uchikata, Takato; Sakamoto, Shigeru; Yokoi, Yasuto; Fukusaki, Eiichiro; Bamba, Takeshi.

In: Journal of Chromatography A, Vol. 1292, 31.05.2013, p. 211-218.

Research output: Contribution to journalArticle

@article{5cf70929b6a9452fb7eecd7b864a10bd,
title = "Development of a lipid profiling system using reverse-phase liquid chromatography coupled to high-resolution mass spectrometry with rapid polarity switching and an automated lipid identification software",
abstract = "Lipidomics requires accurate lipid profiling, which until recently has been challenging at best. In the present study, we developed a practical workflow for high-throughput and exhaustive lipid profiling by combining reverse-phase liquid chromatography coupled to quadrupole orbitrap Fourier transform mass spectrometry, with an automated lipid identification software. This validated method enables highly sensitive and simultaneous analysis of lipids with varying polarities such as glycerophospholipids and sphingophospholipids, by switching the acquisition polarities in mass spectrometry. In addition, it facilitates data-dependent MS2 analysis targeting the lipid molecular species without any influence from other ions by setting the inclusion list, the target m/z list for the product ion scanning. The m/z values of the target lipid molecular species, stored in the database of Lipid Search software, are added to the inclusion list. Moreover, optimizing the identification conditions of the software for the LC/MS system enables high-throughput and accurate identification of lipid molecular species existing in biological samples. Specifically, LC separation is essential for accurate identification of lipid molecular species that possess some fatty acid chains, because it can be difficult to determine fatty acid chain composition of detected molecular species especially in triacylglycerol compounds in direct infusion mass spectrometry. This method has high reproducibility and can be used for structural analysis even for low-abundance compounds. Using this method, over 400 lipid compounds targeted in this research were detected and identified from a sample of mouse plasma. This result indicates that the LC/MS method in the present study enables efficient lipid profiling.",
author = "Takayuki Yamada and Takato Uchikata and Shigeru Sakamoto and Yasuto Yokoi and Eiichiro Fukusaki and Takeshi Bamba",
year = "2013",
month = "5",
day = "31",
doi = "10.1016/j.chroma.2013.01.078",
language = "English",
volume = "1292",
pages = "211--218",
journal = "Journal of Chromatography",
issn = "0021-9673",
publisher = "Elsevier",

}

TY - JOUR

T1 - Development of a lipid profiling system using reverse-phase liquid chromatography coupled to high-resolution mass spectrometry with rapid polarity switching and an automated lipid identification software

AU - Yamada, Takayuki

AU - Uchikata, Takato

AU - Sakamoto, Shigeru

AU - Yokoi, Yasuto

AU - Fukusaki, Eiichiro

AU - Bamba, Takeshi

PY - 2013/5/31

Y1 - 2013/5/31

N2 - Lipidomics requires accurate lipid profiling, which until recently has been challenging at best. In the present study, we developed a practical workflow for high-throughput and exhaustive lipid profiling by combining reverse-phase liquid chromatography coupled to quadrupole orbitrap Fourier transform mass spectrometry, with an automated lipid identification software. This validated method enables highly sensitive and simultaneous analysis of lipids with varying polarities such as glycerophospholipids and sphingophospholipids, by switching the acquisition polarities in mass spectrometry. In addition, it facilitates data-dependent MS2 analysis targeting the lipid molecular species without any influence from other ions by setting the inclusion list, the target m/z list for the product ion scanning. The m/z values of the target lipid molecular species, stored in the database of Lipid Search software, are added to the inclusion list. Moreover, optimizing the identification conditions of the software for the LC/MS system enables high-throughput and accurate identification of lipid molecular species existing in biological samples. Specifically, LC separation is essential for accurate identification of lipid molecular species that possess some fatty acid chains, because it can be difficult to determine fatty acid chain composition of detected molecular species especially in triacylglycerol compounds in direct infusion mass spectrometry. This method has high reproducibility and can be used for structural analysis even for low-abundance compounds. Using this method, over 400 lipid compounds targeted in this research were detected and identified from a sample of mouse plasma. This result indicates that the LC/MS method in the present study enables efficient lipid profiling.

AB - Lipidomics requires accurate lipid profiling, which until recently has been challenging at best. In the present study, we developed a practical workflow for high-throughput and exhaustive lipid profiling by combining reverse-phase liquid chromatography coupled to quadrupole orbitrap Fourier transform mass spectrometry, with an automated lipid identification software. This validated method enables highly sensitive and simultaneous analysis of lipids with varying polarities such as glycerophospholipids and sphingophospholipids, by switching the acquisition polarities in mass spectrometry. In addition, it facilitates data-dependent MS2 analysis targeting the lipid molecular species without any influence from other ions by setting the inclusion list, the target m/z list for the product ion scanning. The m/z values of the target lipid molecular species, stored in the database of Lipid Search software, are added to the inclusion list. Moreover, optimizing the identification conditions of the software for the LC/MS system enables high-throughput and accurate identification of lipid molecular species existing in biological samples. Specifically, LC separation is essential for accurate identification of lipid molecular species that possess some fatty acid chains, because it can be difficult to determine fatty acid chain composition of detected molecular species especially in triacylglycerol compounds in direct infusion mass spectrometry. This method has high reproducibility and can be used for structural analysis even for low-abundance compounds. Using this method, over 400 lipid compounds targeted in this research were detected and identified from a sample of mouse plasma. This result indicates that the LC/MS method in the present study enables efficient lipid profiling.

UR - http://www.scopus.com/inward/record.url?scp=84877135739&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84877135739&partnerID=8YFLogxK

U2 - 10.1016/j.chroma.2013.01.078

DO - 10.1016/j.chroma.2013.01.078

M3 - Article

C2 - 23411146

AN - SCOPUS:84877135739

VL - 1292

SP - 211

EP - 218

JO - Journal of Chromatography

JF - Journal of Chromatography

SN - 0021-9673

ER -