Endocannabinoids are amphiphilic molecules which are synthesized from phospholipids in the nervous system. Together with their G-protein coupled receptors they form the endocannabinoid system. The best studied endocannabinoids are the N-arachidonylethanolamide (anandamide) and the 2-arachidonoylglycerol (2-AG). They consist of an amide or an ester head, and linked with an arachidonyl chain. The physiological activities of Endocannabinoids mainly occur by binding to cannabinoid receptors or TRP channels. Moreover, it has been demonstrated that the endocannabinoids are involved in many membrane proteins in the presence of antagonists of their classical receptors, indicating the endocannabinoids can mediate physiological activities in a receptor-independent way.
Figure 1. Structure of endocannabinoids anandamide and 2-AG
The endocannabinoids have been shown to regulate a wide range of physiological and cognitive processes including fertility, pregnancy, during pre- and postnatal development, motor control, appetite, food intake, pain-sensation, mood, memory acquisition and learning. In humans, exercise has been shown to induce the increase of the endocannabinoids levels in plasma, mainly because endocannabinoids can effectively cross the blood–brain barrier, and contribute to the development of exercise-induced euphoria in humans. Disorders of the endocannabinoid system have been related to several neuro-inflammatory diseases such as Alzheimer, Parkinson, Huntington and Multiple Sclerosis.
Because of their low abundance and structural variability, the quantification of endogenous endocannabinoid is challenging. LC–MS/MS platform enable simultaneous relative and absolute quantification of multiple metabolites in this pathway. The results are widely used for investigating the relative changes in metabolite concentrations, and potential biomarker discovery. Creative Proteomics has established sensitive, reliable, and accurate HPLC-MS/MS method for quantification of endocannabinoid metabolites.
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