Analytical and Bioanalytical Chemistry

, Volume 396, Issue 7, pp 2415–2423

Identification of 48 homologues of phosphatidylethanol in blood by LC-ESI-MS/MS

  • H. Gnann
  • C. Engelmann
  • G. Skopp
  • M. Winkler
  • V. Auwärter
  • S. Dresen
  • N. Ferreirós
  • F. M. Wurst
  • W. Weinmann
Original Paper

DOI: 10.1007/s00216-010-3458-5

Cite this article as:
Gnann, H., Engelmann, C., Skopp, G. et al. Anal Bioanal Chem (2010) 396: 2415. doi:10.1007/s00216-010-3458-5

Abstract

Phosphatidylethanol (PEth) is an abnormal phospholipid carrying two fatty acid chains. It is only formed in the presence of ethanol via the action of phospholipase D (PLD). Its use as a biomarker for alcohol consumption is currently under investigation. Previous methods for the analysis of PEth included high-performance liquid chromatography (HPLC) coupled to an evaporative light scattering detector (ELSD), which is unspecific for the different homologues—improved methods are now based on time of flight mass spectrometry (TOF-MS) and tandem mass spectrometry (MS/MS). The intention of this work was to identify as many homologues of PEth as possible. A screening procedure using multiple-reaction monitoring (MRM) for the identified homologues has subsequently been established. For our investigations, autopsy blood samples collected from heavy drinkers were used. Phosphatidylpropanol 16:0/18:1 (internal standard) was added to the blood samples prior to liquid–liquid extraction using borate buffer (pH 9), 2-propanol and n-hexane. After evaporation, the samples were redissolved in the mobile phase and injected into the LC-MS/MS system. Compounds were separated on a Luna Phenyl Hexyl column (50 mm × 2 mm, 3 µm) by gradient elution, using 2 mM ammonium acetate and methanol/acetone (95/5; v/v). A total of 48 homologues of PEth could be identified by using precursor ion and enhanced product ion scans (EPI).

Keywords

AlcoholPhosphatidylethanolPhospholipidsHomologuesLC-MS/MSBiomarker

Supplementary material

216_2010_3458_MOESM1_ESM.pdf (875 kb)
ESM 1(PDF 875 kb)

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • H. Gnann
    • 1
  • C. Engelmann
    • 1
  • G. Skopp
    • 2
  • M. Winkler
    • 3
  • V. Auwärter
    • 1
  • S. Dresen
    • 1
  • N. Ferreirós
    • 1
  • F. M. Wurst
    • 4
  • W. Weinmann
    • 1
  1. 1.Institute of Legal MedicineUniversity Medical CentreFreiburgGermany
  2. 2.Institute of Legal Medicine and Traffic MedicineUniversity HospitalHeidelbergGermany
  3. 3.Institute of Legal MedicineUniversity of UlmUlmGermany
  4. 4.Department of Psychiatry and Psychotherapy II, Christian Doppler ClinicParacelsus Medical UniversitySalzburgAustria