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Peripheral endocannabinoid microdialysis: in vitro characterization and proof-of-concept in human subjects

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Abstract

In vivo endocannabinoid (EC) microdialysis has only seldom been performed, mostly in rodent brain tissue. Low solubility in aqueous media, adsorption to surfaces, and instability with co-present human serum albumin (HSA) are the major obstacles in EC microdialysis. The addition of hydroxypropyl-ß-cyclodextrine (HPCD) to the perfusion fluid has been previously described to facilitate lipid microdialysis, but the general biophysical properties of HPCD, especially with respect to peripheral EC microdialysis, have not been described before. We report on the characterization of EC microdialysis using an in vitro system using Ringer’s solution with 10% HPCD as the perfusion fluid and with fatty acid-free HSA as the matrix fluid. The endocannabinoids anandamide (AEA) and 2-arachidonoyl glycerol (2AG) were measured using LC-MS/MS. AEA was stable in the perfusion and matrix fluids, whereas 2AG was only stable in the perfusion fluid. In the matrix fluid, 2AG underwent rapid isomerization to 1-arachidonoyl glycerol. A relative recovery of 3.5% for AEA was found with 10% HPCD in the perfusion fluid and a flow rate of 1 μL/min. For 2AG, a similar relative recovery of 3.5% was estimated. Since 2AG was found unstable in the matrix fluid, a reliable calculation of the relative recovery rates was not possible. Delivery and recovery experiments revealed unequal inward and outward EC transport across the microdialysis membrane. Contrary to usual microdialysis findings, we observed increasing recovery rates for AEA with increasing flow rates. Long equilibration times of several hours were necessary to obtain constant relative recovery rates. In a proof-of-concept study in humans, we collected AEA from subcutaneous abdominal adipose tissue employing the described methodology. Our study suggests that the microdialysis technique is not suitable for the exact quantification of tissue EC concentrations, but it allows for their rough estimation.

Microdialysis of peripheral endocannabinoids, especially anandamide (AEA).

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Acknowledgments

S.B. was supported by Alexander von Humboldt Foundation. This work was supported by the Commission of 581 the European Communities (Collaborative Project ADAPT, contract 582 number HEALTH-F2-2008-201100). Support and helpful advice from Loren H. Parsons, Committee on the Neurobiology of Addictive Disorders, The Scripps Research Institute, La Jolla, California, USA and from C. Kloft and F. Simmel at the Institute of Clinical Pharmacy at University Halle-Wittenberg, Germany, are greatly appreciated.

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  1. Institute for Clinical Pharmacology, Hannover Medical School, Carl-Neuberg-Straße 1, 30625, Hannover, Germany

    Alexander A. Zoerner, Christin Rakers, Stefan Engeli, Sandor Batkai, Marcus May, Jens Jordan & Dimitrios Tsikas

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  1. Alexander A. Zoerner
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  2. Christin Rakers
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  3. Stefan Engeli
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  4. Sandor Batkai
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  7. Dimitrios Tsikas
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Correspondence to Alexander A. Zoerner.

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Zoerner, A.A., Rakers, C., Engeli, S. et al. Peripheral endocannabinoid microdialysis: in vitro characterization and proof-of-concept in human subjects. Anal Bioanal Chem 402, 2727–2735 (2012). https://doi.org/10.1007/s00216-012-5729-9

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  • DOI: https://doi.org/10.1007/s00216-012-5729-9

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