Electrospray ionization mass spectrometric method for the determination of cannabinoid precursors: N-acylethanolamine phospholipids (NAPEs).
N-Acylethanolamine phospholipids (NAPEs) serve as endogenous precursors of N-acylethanolamines (NAEs), e.g. N-arachidonoylethanolamine (anandamide) and N-palmitoylethanolamine that are endogenous ligands of cannabinoid receptors. Under physiological conditions, NAPE is found in very low concentrations in mammalian tissue (3-12 nmol g(-1)). However, pathophysiological conditions may increase the endogenous NAPE levels, which again may cause an increase in endocannabinoid concentrations. This paper presents a simple and selective method for the determination of NAPE standards using negative electrospray ionization mass spectrometry (ESI-MS). The procedure provides complete positioning of all acyl and alkenyl groups contained in each NAPE species. The calibration curve for standard NAPE was linear over the range 100 fmol-50 pmol (0.1-50 ng) per injection. The lower limit of detection (signal-to-noise ratio of 3) was 100 fmol, implying that this method is superior to previous methods for the determination of NAPE. These results suggest that this ESI-MS method can be used to identify and quantify NAPE species in mammalian tissues and provide information on the corresponding NAEs to be released from the endogenous NAPE pool.
Department of Pharmacology, Royal Danish School of Pharmacy, 2 Universitetsparken, DK-2100 Copenhagen, Denmark. hhh@mail.dfh.dk
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D000818:Animals; D001921:Brain / Q000378:metabolism; D063385:Cannabinoid Receptor Modulators; D002186:Cannabinoids / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D004983:Ethanolamines / Q000032:analysis / Q000737:chemistry*; D008024:Ligands; D013058:Mass Spectrometry / Q000379:methods; D015394:Molecular Structure; D010743:Phospholipids / Q000032:analysis / Q000737:chemistry*; D043882:Receptors, Cannabinoid; D011955:Receptors, Drug / Q000378:metabolism
D063385:Cannabinoid Receptor Modulators; D002186:Cannabinoids; D004983:Ethanolamines; D008024:Ligands; C022203:N-acylethanolamines; D010743:Phospholipids; D043882:Receptors, Cannabinoid; D011955:Receptors, Drug