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

Analytical and Bioanalytical Chemistry volume 396, pages 2415–2423 (2010)Cite this article

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).

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Acknowledgements

N. Ferreirós thanks the government of Basque Country for her postdoctoral grant.

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Authors and Affiliations

  1. Institute of Legal Medicine, University Medical Centre, Albertstr. 9, 79104, Freiburg, Germany

    H. Gnann, C. Engelmann, V. Auwärter, S. Dresen, N. Ferreirós & W. Weinmann

  2. Institute of Legal Medicine and Traffic Medicine, University Hospital, 69115, Heidelberg, Germany

    G. Skopp

  3. Institute of Legal Medicine, University of Ulm, 89075, Ulm, Germany

    M. Winkler

  4. Department of Psychiatry and Psychotherapy II, Christian Doppler Clinic, Paracelsus Medical University, 5020, Salzburg, Austria

    F. M. Wurst

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  1. H. Gnann
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  2. C. Engelmann
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  4. M. Winkler
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  5. V. Auwärter
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  6. S. Dresen
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  7. N. Ferreirós
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  8. F. M. Wurst
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Correspondence to W. Weinmann.

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Gnann, H., Engelmann, C., Skopp, G. et al. Identification of 48 homologues of phosphatidylethanol in blood by LC-ESI-MS/MS. Anal Bioanal Chem 396, 2415–2423 (2010). https://doi.org/10.1007/s00216-010-3458-5

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  • DOI: https://doi.org/10.1007/s00216-010-3458-5

Keywords

  • Alcohol
  • Phosphatidylethanol
  • Phospholipids
  • Homologues
  • LC-MS/MS
  • Biomarker