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Analytical and Bioanalytical Chemistry

, Volume 407, Issue 6, pp 1781–1787 | Cite as

A simple method for simultaneous determination of N-arachidonoylethanolamine, N-oleoylethanolamine, N-palmitoylethanolamine and 2-arachidonoylglycerol in human cells

  • Igor IvanovEmail author
  • Philipp Borchert
  • Burkhard Hinz
Note

Abstract

The endocannabinoid system has been considered as a target for pharmacological intervention. Accordingly, inhibition of fatty acid amide hydrolase (FAAH), a degrading enzyme of the endocannabinoids N-arachidonoylethanolamine (anandamide; AEA) and 2-arachidonoylglycerol (2-AG) as well as of the endocannabinoid-like substances N-oleoylethanolamine (OEA) and N-palmitoylethanolamine (PEA), can cause augmented endogenous cannabinoid tone. Using liquid chromatography coupled with positive electrospray ionisation mass spectrometry, we herein describe a method to simultaneously quantify levels of AEA, OEA, PEA and 2-AG in cultured cells. The procedure was developed according to the FDA guidelines for bioanalytical methods validation. The limits of quantification (LOQs) were 0.05 pmol for AEA, 0.09 pmol for OEA, 0.10 pmol for PEA and 0.80 pmol for 2-AG when molecular ion monitoring was used. In H460 human lung carcinoma cells, basal levels of all four analytes ranged between 2 and 17 pmol mg−1 protein with PEA showing the lowest and OEA the highest concentrations. Endocannabinoid levels observed in mesenchymal stem cells were of the same order of magnitude when compared to those in H460 human lung carcinoma cells.

Keywords

LC–MS/MS Endocannabinoids Endocannabinoid-like substances H460 Lung carcinoma cells Mesenchymal stem cells 

Notes

Acknowledgments

The authors wish to thank Ms Y. Wollank for preparing mesenchymal stem cells and Ms A.M. Berg for critical reading of the manuscript.

Supplementary material

216_2014_8384_MOESM1_ESM.pdf (36 kb)
ESM 1 (PDF 35 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  1. 1.Institute of Toxicology and PharmacologyUniversity of RostockRostockGermany

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