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Determination of the two major endocannabinoids in human plasma by μ-SPE followed by HPLC-MS/MS

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Abstract

Endocannabinoids (ECs) are endogenous compounds that interact with type-1 and type-2 cannabinoid receptors (CB1 and CB2), as well as non-cannabinoid receptors. The multitude of roles attributed to ECs makes them an emerging target of pharmacotherapy for a number of disparate diseases. Here a high-throughput bioanalytical method based on micro SPE (μ-SPE) followed by LC-MS/MS analysis for the simultaneous determination of the two major endocannabinoids 2-arachidonoylglycerol (2-AG) and N-arachidonoylethanolamine (anandamide, AEA) in human plasma is presented. The chromatographic conditions obtained with the fused-core column allowed a good separation in 10 min also of the AG isomers. A very simple and reliable extraction has been optimised by means of C18-modified tips: it requires only 100 μL of plasma and allows the use of minimal volumes of organic solvent. The present method allows a rapid and effective clean-up, which also minimises the isomerisation of 2-AG. The whole procedure has been validated following the FDA guidelines for bioanalytical methods validation: the satisfactory recovery values, the negligible matrix effect and the good values of accuracy and reproducibility make it a simple and high-throughput analytical tool for clinical and biochemical studies on endocannabinoid signaling in humans.

Determination of the two major endocannabinoids in human plasma by μ-SPE followed by HPLC-MS/MS

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Aknowledgements

Financial support from Fondazione TERCAS (grant 2009-2012 to MM) is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Food Sciences, University of Teramo, 64100, Teramo, Italy

    Manuel Sergi & Dario Compagnone

  2. Department of Biomedical Sciences, University of Teramo, 64100, Teramo, Italy

    Natalia Battista & Mauro Maccarrone

  3. European Center for Brain Research (CERC)/Santa Lucia Foundation, 00164, Rome, Italy

    Natalia Battista & Mauro Maccarrone

  4. Department of Chemistry, Sapienza University of Rome, 00185, Rome, Italy

    Camilla Montesano & Roberta Curini

Authors
  1. Manuel Sergi
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  2. Natalia Battista
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  3. Camilla Montesano
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  4. Roberta Curini
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  5. Mauro Maccarrone
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  6. Dario Compagnone
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Corresponding author

Correspondence to Dario Compagnone.

Additional information

Published in the special issue Analytical Science in Italy with guest editor Aldo Roda

Mauro Maccarrone and Dario Compagnone are equally senior authors.

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Sergi, M., Battista, N., Montesano, C. et al. Determination of the two major endocannabinoids in human plasma by μ-SPE followed by HPLC-MS/MS. Anal Bioanal Chem 405, 785–793 (2013). https://doi.org/10.1007/s00216-012-6273-3

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  • DOI: https://doi.org/10.1007/s00216-012-6273-3

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