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

Manuel Sergi, Natalia Battista, Camilla Montesano, Roberta Curini, Mauro MacCarrone, Dario Compagnone

Research output: Contribution to journalArticlepeer-review


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. [Figure not available: see fulltext.]

Original languageEnglish
Pages (from-to)785-793
Number of pages9
JournalAnalytical and Bioanalytical Chemistry
Issue number2-3
Publication statusPublished - 2013


  • 2-Arachidonoylglycerol (2-AG)
  • Arachidonoylethanolamine (anandamide, AEA)
  • Endocannabinoids
  • LC-MS/MS
  • Plasma

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry


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