Skip to main content

Advertisement

Log in

LC-MS/MS analysis of phosphatidylethanol in dried blood spots versus conventional blood specimens

  • Technical Note
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Phosphatidylethanol (PEth), which is formed extrahepatically by the action of phospholipase D on phosphatidylcholine in the presence of ethanol, has been suggested as a promising marker of alcohol misuse. Analysis of dried blood spots (DBS) is particularly advantageous for the determination of delicate analytes such as PEth. Therefore, measurement of PEth species (18:1/18:1, 16:0/18:1) in DBS versus whole blood was performed to ascertain whether respective results are directly comparable. Samples were obtained from subjects (n = 40) undergoing alcohol detoxification treatment. Analysis involved liquid–liquid extraction from both, DBS and whole blood (100 μL, respectively), with phosphatidylpropanol as the internal standard. Extracts were subjected to LC gradient separation using multiple reaction monitoring of deprotonated molecules. Results from measurements of corresponding DBS and whole blood specimens were compared by estimating the respective mean values and by a Bland and Altman analysis. Concentrations of PEth 18:1/18:1 ranged from 46.1 to 3,360 ng/mL in whole blood (mean, 461.7 ng/mL) and from 35.8 to 3,360 ng/mL in DBS (mean, 457.6 ng/mL); for PEth 16:0/18:1, concentrations were from 900 to 213,000 ng/mL (mean, 23,375 ng/mL) and 922–213,000 ng/mL (mean, 23,470 ng/mL) in blood and DBS, respectively. Estimated mean differences were −4.3 ng/mL for PEth 18:1/18:1 and 95.8 ng/mL for PEth 16:0/18:1. The Bland–Altman plot of both PEth species showed that the variation around the mean difference was similar all through the range of measured values and that all differences except one were within the limits of agreement. It could be shown that the determination of PEth species in DBS is as reliable as in whole blood samples. This assay may facilitate monitoring of alcohol misuse.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Aradottir S, Seidl S, Wurst FM, Jonsson BA, Alling C (2004) Phosphatidylethanol in human organs and blood: a study on autopsy material and influences by storage conditions. Alcohol Clin Exp Res 28:1718–1723

    Article  CAS  Google Scholar 

  2. Gnann H, Weinmann W, Engelmann C, Wurst FM, Skopp G, Winkler M et al (2009) Selective detection of phosphatidylethanol homologues in blood as biomarkers for alcohol consumption by LC-ESI-MS/MS. J Mass Spectrom 44:1293–1299

    Article  CAS  Google Scholar 

  3. Aradottir S, Olsson BL (2005) Methodological modifications on quantification of phosphatidylethanol in blood from humans abusing alcohol, using high-performance liquid chromatography and evaporative light scattering detection. BMC Biochem 6:18

    Article  Google Scholar 

  4. Skopp G, Potsch L (2001) Detection of cocaine in blood stains. Arch Kriminol 207:81–88

    CAS  Google Scholar 

  5. Garcia Boy R, Henseler J, Mattern R, Skopp G (2008) Determination of morphine and 6-acetylmorphine in blood with use of dried blood spots. Ther Drug Monit 30:733–739

    Article  CAS  Google Scholar 

  6. Services USDoHaH, Administration FaD, (CDER) CfDEaR, (CVM) CfVM (2001) Guidance for Industry—Bioanalytical Method Validation

  7. Mantha S, Roizen MF, Fleisher LA, Thisted R, Foss J (2000) Comparing methods of clinical measurement: reporting standards for Bland and Altman analysis. Anesth Analg 90:593–602

    Article  CAS  Google Scholar 

  8. Nalesso A, Viel G, Cecchetto G, Frison G, Ferrara SD (2010) Analysis of the alcohol biomarker phosphatidylethanol by NACE with on-line ESI-MS. Electrophoresis 31:1227–1233

    Article  CAS  Google Scholar 

  9. Leidl K, Liebisch G, Richter D, Schmitz G (2008) Mass spectrometric analysis of lipid species of human circulating blood cells. Biochim Biophys Acta 1781:655–664

    CAS  Google Scholar 

  10. Jakobik V, Burus I, Decsi T (2009) Fatty acid composition of erythrocyte membrane lipids in healthy subjects from birth to young adulthood. Eur J Pediatr 168:141–147

    Article  CAS  Google Scholar 

  11. Mei JV, Alexander JR, Adam BW, Hannon WH (2001) Use of filter paper for the collection and analysis of human whole blood specimens. J Nutr 131:1631–1637

    Google Scholar 

  12. Parker SP, Cubitt WD (1999) The use of the dried blood spot sample in epidemiological studies. J Clin Pathol 52:633–639

    Article  CAS  Google Scholar 

  13. Gnann H, Engelmann C, Skopp G, Winkler M, Auwarter V, Dresen S et al (2010) Identification of 48 homologues of phosphatidylethanol in blood by LC-ESI-MS/MS. Anal Bioanal Chem 396:2415–2423

    Article  CAS  Google Scholar 

Download references

Acknowledgment

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

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Andrea Faller.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Faller, A., Richter, B., Kluge, M. et al. LC-MS/MS analysis of phosphatidylethanol in dried blood spots versus conventional blood specimens. Anal Bioanal Chem 401, 1163–1166 (2011). https://doi.org/10.1007/s00216-011-5221-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-011-5221-y

Keywords

Navigation