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Analytical and Bioanalytical Chemistry

, Volume 400, Issue 7, pp 1923–1931 | Cite as

Liquid chromatography–mass spectrometric determination of plasmalogens in human plasma

  • Shu-Ping Hui
  • Hitoshi ChibaEmail author
  • Takao Kurosawa
Original Paper

Abstract

A new liquid chromatography–mass spectrometry (LC/MS) method has been developed for the quantitative analysis of plasmalogens in human plasma using a nonendogenous plasmalogen (1-O-1′-(Z)-tricosenyl-2-oleoyl-rac-glycero-3-phosphocholine, PLS 23:0/18:1) as an internal standard. 1-O-1′-(Z)-Tricosenyl glyceryl ether was prepared by reacting lithioalkoxyallyl with 1-iodoeicosane as the key intermediate in the formation of PLS 23:0/18:1. In LC/MS analyses, PLS 23:0/18:1 generated significant fragment ions in positive and negative modes. In positive ion mode, the [M+H]+ of PLS 23:0/18:1 yielded unique fragments with cleavages at the sn-1 and sn-2 positions of the glycerol backbone. In negative ion mode, the [M+CH3COO] of PLS 23:0/18:1 resulted in characteristic fragmentation at the sn-2 and sn-3 positions. 1-O-1′-(Z)-Hexadecenyl-2-linoleoyl-rac-glycero-3-phosphocholine (PLS 16:0/18:2) and 2-arachidonoyl-O-1′-(Z)-hexadecenyl-rac-glycero-3-phosphocholine (PLS 16:0/20:4) were chemically synthesized as PLS 23:0/18:1. The calibration curves obtained for PLS 16:0/18:2 and PLS 16:0/20:4 were linear throughout the calibration range (0.04–1.60 pmol). The LOD (S/N = 5:1) was 0.008 pmol and the LOQ (S/N = 6:1) was 0.01 pmol for both PLS 16:0/18:2 and PLS 16:0/20:4. Plasma concentrations of PLS 16:0/18:2 and PLS 16:0/20:4 were 4.0 ± 1.3 μM and 3.5 ± 1.2 μM (mean ± SD), respectively, in five healthy volunteers.

Figure

SRM chromatograms were obtained by the quantitative LC/MS system in positive mode: (A) synthetic PLS 16:0/18:2, (B) synthetic PLS 16:0/20:4, (C) IS, (A') plasma extract (precursor ion: m/z 742.6), (B') plasma extract (precursorion: m/z 766.6), (C') plasma extract after addition of the IS.

Keywords

Liquid chromatography/electrospray ionization mass spectrometry LC/MS Plasmalogen Analysis Synthesis 

Abbreviations

ESI-MS

Electrospray ionization mass spectrometry

IS

Internal standard

LC/MS

Liquid chromatography/mass spectrometry

LOD

Limit of detection

LOQ

Limit of quantification

PLS 16:0/18:2

1-O-1′-(Z)-hexadecenyl-2-linoleoyl-rac-glycero-3-phosphocholine

PLS 16:0/20:4

2-Arachidonoyl-O-1′-(Z)-hexadecenyl-rac-glycero-3-phosphocholine

PLS 23:0/18:1

1-O-1′-(Z)-tricosenyl-2-oleoyl-rac-glycero-3-phosphocholine

SEMCl

2-(Chloromethoxy) ethyltrimethylsilane

SRM

Selected reaction monitoring

TBAF

Tetrabutylammonium fluoride

TBSCl

Tert-butyldimethylchlorosilane

THF

Tetrahydrofuran

Notes

Acknowledgments

This study was supported by Sapporo Biocluster “Bio-S”, The Regional Innovation Cluster Program, The Ministry of Education, Culture, Sports, Science and Technology, Japan.

Supplementary material

216_2011_4921_MOESM1_ESM.pdf (57 kb)
ESM 1 (PDF 57 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Faculty of Health Sciences, Medical Laboratory SciencesHokkaido UniversitySapporoJapan
  2. 2.Faculty of Pharmaceutical SciencesHealth Sciences University of HokkaidoHokkaidoJapan
  3. 3.Faculty of Health SciencesHokkaido University Graduate School of Health SciencesSapporoJapan

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