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International Journal of Legal Medicine

, Volume 127, Issue 3, pp 603–610 | Cite as

Stability of phosphatidylethanol species in spiked and authentic whole blood and matching dried blood spots

  • Andrea Faller
  • Barbara Richter
  • Matthias Kluge
  • Patrick Koenig
  • H. K. Seitz
  • Gisela SkoppEmail author
Original Article

Abstract

Background

Phosphatidylethanol (PEth) is currently under investigation as a highly sensitive and specific marker of alcohol misuse. As its stability in blood samples has not systematically been investigated, a study was performed to determine the stability of major PEth species in spiked and authentic whole blood and also in matching dried blood spots (DBS) at different conditions.

Methods

To PEth-free blood from teetotalers, low and high concentrations of two major PEth (18:1/18:1 and 16:0/18:1) species were added chosen on the basis of concentrations determined from authentic samples which were collected from the subjects undergoing alcohol detoxification treatment. Effects of sampling (EDTA or heparinized tubes), temperature, and time (≤30 days) were investigated. Processed samples (two at each condition, respectively) were subjected to LC gradient separation using multiple reaction monitoring. Stability was assessed using the critical difference or a periodic analysis result that was within 15 % of the initial concentration. Reaction kinetics of degradation was investigated with rate constants being checked for an Arrhenius relationship.

Results

PEth was stable in dried blood spot (DBS) stored either at room temperature or frozen, whereas it was not stable in whole blood except in samples stored at −80 °C. Activation energies increased in the following order: spiked heparinized blood < spiked EDTA blood < authentic EDTA blood.

Conclusions

PEth is a labile analyte which is predominantly degraded by hydrolysis. Only at −80 °C, stability in whole blood can be ascertained, and analysis should be performed within 30 days. EDTA should be preferred over heparin as an additive. DBS is able to stabilize PEth thus partly resolving pre-analytical difficulties of PEth measurement.

Keywords

Stability Dried blood spots Whole blood Phosphatidylethanol LC-MS/MS 

Notes

Acknowledgments

This project was funded by the Ministry of Education of Baden-Württemberg, Germany.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Andrea Faller
    • 1
  • Barbara Richter
    • 2
  • Matthias Kluge
    • 2
  • Patrick Koenig
    • 3
  • H. K. Seitz
    • 3
  • Gisela Skopp
    • 1
    Email author
  1. 1.Institute of Legal and Traffic MedicineUniversity Hospital HeidelbergHeidelbergGermany
  2. 2.Psychiatric Center NordbadenWieslochGermany
  3. 3.Salem Medical CenterHeidelbergGermany

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