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Analytical and Bioanalytical Chemistry

, Volume 405, Issue 14, pp 4981–4987 | Cite as

Analysis of triacylglycerol hydroperoxides in human lipoproteins by Orbitrap mass spectrometer

  • Shu-Ping HuiEmail author
  • Toshihiro Sakurai
  • Seiji Takeda
  • Shigeki Jin
  • Hirotoshi Fuda
  • Takao Kurosawa
  • Hitoshi ChibaEmail author
Note

Abstract

Herein, we represent a simple method for the detection and characterization of molecular species of triacylglycerol monohydroperoxides (TGOOH) in biological samples by use of reversed-phase liquid chromatography with a LTQ Orbitrap XL mass spectrometer (LC/LTQ Orbitrap) via an electrospray ionization source. Data were acquired using high-resolution, high-mass accuracy in Fourier-transform mode. Platform performance, related to the identification of TGOOH in human lipoproteins and plasma, was estimated using extracted ion chromatograms with mass tolerance windows of 5 ppm. Native low-density lipoproteins (nLDL) and native high-density lipoproteins (nHDL) from a healthy donor were oxidized by CuSO4 to generate oxidized LDL (oxLDL) and oxidized HDL (oxHDL). No TGOOH molecular species were detected in the nLDL and nHDL, whereas 11 species of TGOOH molecules were detected in the oxLDL and oxHDL. In positive-ion mode, TGOOH was found as [M + NH4]+. In negative-ion mode, TGOOH was observed as [M + CH3COO]. TGOOH was more easily ionized in positive-ion mode than in negative-ion mode. The LC/LTQ Orbitrap method was applied to human plasma and three molecular species of TGOOH were detected. The limit of detection is 0.1 pmol (S/N = 10:1) for each synthesized TGOOH.

Figure

Analysis of triacylglycerol hydroperoxides in human lipoproteins by Orbitrap mass spectrometer

Keywords

Triacylglycerol hydroperoxides TGOOH Orbitrap Liquid chromatography/electrospray ionization mass spectrometry LC/MS Triglyceride 

Abbreviations

EIC

Extracted ion chromatogram(s)

HPLC

High-performance liquid chromatography

LC/LTQ Orbitrap

Reversed-phase liquid chromatography with a LTQ Orbitrap XL mass spectrometer

LC/MS

Liquid chromatography/mass spectrometry

nHDL

Native high-density lipoproteins

nLDL

Native low-density lipoproteins

oxHDL

Oxidized high-density lipoproteins

oxLDL

Oxidized low-density lipoproteins

RT

Retention time

TGOOH

Triacylglycerol monohydroperoxide(s)

TGOOH 18:1/18:1/16:0

1,2-Dioleoyl-3-palmitoylglycerol monohydroperoxide(s)

TGOOH 18:1/18:1/18:1

Triolein monohydroperoxide (s)

TGOOH 18:1/18:2/16:0

1-Oleoyl-2-linoleoyl-3-palmitoylglycerol monohydroperoxide(s)

TIC

Total ion current chromatogram(s)

Notes

Acknowledgments

This study was supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science, by the Regional Innovation Strategy Support Program, Sapporo Health Innovation “Smart-H”, of the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Shu-Ping Hui
    • 1
    Email author
  • Toshihiro Sakurai
    • 1
  • Seiji Takeda
    • 1
  • Shigeki Jin
    • 1
  • Hirotoshi Fuda
    • 1
  • Takao Kurosawa
    • 2
  • Hitoshi Chiba
    • 1
    Email author
  1. 1.Faculty of Health SciencesHokkaido UniversitySapporoJapan
  2. 2.Faculty of Pharmaceutical SciencesHealth Sciences University of HokkaidoHokkaidoJapan

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