title: Determination of the two major endocannabinoids in human plasma by μ-SPE followed by HPLC-MS/MS.
abstract: Endocannabinoids (ECs) are endogenous compounds that interact with type-1 and type-2 cannabinoid receptors (CB(1) and CB(2)), 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.
authors: Department of Food Sciences, University of Teramo, Teramo, Italy.
authors: Manuel
authors: M
authors: Sergi
authors: Natalia
authors: N
authors: Battista
authors: Camilla
authors: C
authors: Montesano
authors: Roberta
authors: R
authors: Curini
authors: Mauro
authors: M
authors: Maccarrone
authors: Dario
authors: D
authors: Compagnone
mesh_terms: D001095:Arachidonic Acids / Q000097:blood* / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D063388:Endocannabinoids / Q000097:blood* / Q000302:isolation & purification*; D005989:Glycerides / Q000097:blood* / Q000302:isolation & purification*; D006801:Humans; D053284:Polyunsaturated Alkamides; D052617:Solid Phase Microextraction / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001095:Arachidonic Acids; D063388:Endocannabinoids; D005989:Glycerides; D053284:Polyunsaturated Alkamides; C094503:glyceryl 2-arachidonate; C078814:anandamide