Endocannabinoids (ECs) are endogenous compoundsthat interact with type-1 and type-2 cannabinoidreceptors (CB1 and CB2), as well as non-cannabinoid receptors.The multitude of roles attributed to ECs makes them anemerging target of pharmacotherapy for a number of disparatediseases. Here a high-throughput bioanalytical methodbased on micro SPE (μ-SPE) followed by LC-MS/MS analysisfor the simultaneous determination of the two majorendocannabinoids 2-arachidonoylglycerol (2-AG) and Narachidonoylethanolamine(anandamide, AEA) in humanplasma is presented. The chromatographic conditionsobtained with the fused-core column allowed a good separationin 10 min also of the AG isomers. A very simple andreliable extraction has been optimised by means of C18-modified tips: it requires only 100 μL of plasma and allowsthe use of minimal volumes of organic solvent. The present method allows a rapid and effective clean-up, which alsominimises the isomerisation of 2-AG. The whole procedurehas been validated following the FDA guidelines for bioanalyticalmethods validation: the satisfactory recovery values,the negligible matrix effect and the good values ofaccuracy and reproducibility make it a simple and highthroughputanalytical tool for clinical and biochemical studieson endocannabinoid signaling in humans.[...]
Determination of the two major endocannabinoids in human plasma by μ-SPE followed by HPLC-MS/MS
SERGI, Manuel;BATTISTA, Natalia;COMPAGNONE, DARIO
2013-01-01
Abstract
Endocannabinoids (ECs) are endogenous compoundsthat interact with type-1 and type-2 cannabinoidreceptors (CB1 and CB2), as well as non-cannabinoid receptors.The multitude of roles attributed to ECs makes them anemerging target of pharmacotherapy for a number of disparatediseases. Here a high-throughput bioanalytical methodbased on micro SPE (μ-SPE) followed by LC-MS/MS analysisfor the simultaneous determination of the two majorendocannabinoids 2-arachidonoylglycerol (2-AG) and Narachidonoylethanolamine(anandamide, AEA) in humanplasma is presented. The chromatographic conditionsobtained with the fused-core column allowed a good separationin 10 min also of the AG isomers. A very simple andreliable extraction has been optimised by means of C18-modified tips: it requires only 100 μL of plasma and allowsthe use of minimal volumes of organic solvent. The present method allows a rapid and effective clean-up, which alsominimises the isomerisation of 2-AG. The whole procedurehas been validated following the FDA guidelines for bioanalyticalmethods validation: the satisfactory recovery values,the negligible matrix effect and the good values ofaccuracy and reproducibility make it a simple and highthroughputanalytical tool for clinical and biochemical studieson endocannabinoid signaling in humans.[...]I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
