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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.


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1437-4331
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Release of anandamide from blood cells

Michael Vogeser1 / Daniela Hauer2 / Shahnaz Christina Azad3 / Erasmus Huber4 / Martin Storr5 / Gustav Schelling6

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Corresponding author: PD Dr. med. Michael Vogeser, Institute of Clinical Chemistry, Hospital of the University of Munich, Marchioninistr. 15, 81377 Munich, Germany Phone: +49-89-70953221, Fax: +49-89-70953240

Citation Information: Clinical Chemical Laboratory Medicine. Volume 44, Issue 4, Pages 488–491, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: 10.1515/CCLM.2006.065, April 2006

Publication History

Received:
September 5, 2005
Accepted:
January 3, 2006
Published Online:
2006-04-07

Abstract

Background: Endogenous ligands of cannabinoid receptors (endocannabinoids), in particular anandamide (arachidonylethanolamide), have been recognized as being of crucial importance in a variety of physiological functions. Plasma concentrations of anandamide have been measured in a number of investigations; however, discrepant data on “normal” anandamide plasma concentrations were reported. Since this might be caused by pre-analytical variables, we investigated the impact of different sample handling conditions on measured plasma anandamide concentrations.

Methods: Blood samples were taken from healthy volunteers in EDTA- or heparin-containing tubes; whole blood samples were kept at +4°C, room temperature, or 37°C, respectively, for up to 120 min before obtaining plasma by centrifugation. Plasma anandamide concentrations were measured by an isotope-dilution liquid chromatography tandem mass spectrometry (LC-MS/MS) method.

Results: A marked time- and temperature-dependent increase in plasma anandamide concentrations ex vivo was observed in both EDTA- and heparin-containing tubes. Mean anandamide concentrations approximately doubled when EDTA samples were kept at 4°C for 60 min before centrifugation [immediately centrifuged, 1.3 μg/L (SD 0.3 μg/L); 2.8 μg/L (SD 0.5 μg/L) after storage for 60 min; n=12). After storage of heparinized whole-blood samples for 120 min at 37°C, a mean plasma anandamide concentration of 11.9 μg/L (SD 1.8 μg/L) was found. In cell-free plasma, no increase in anandamide concentrations was found.

Conclusion: Anandamide is released from blood cells ex vivo at a very high rate; therefore, strictly standardized pre-analytical protocols have to be applied for plasma anandamide determination.

Keywords: anandamide (arachidonylethanolamide); liquid chromatography-tandem mass spectrometry; preanalytical period

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