ANALYSIS OF ANANDAMIDE, AN ENDOGENOUS CANNABINOID SUBSTANCE, AND OF OTHER NATURAL N-ACYLETHANOLAMINES

Recent reports have suggested that N-arachidonoylethanolamine (anandam ide) acts as an endogenous ligand for cannabinoid receptors in mammali an brain. Here we describe methods for the extraction, purification an d analysis of anandamide and related N-fatty acyl-ethanolamines (NAEs) . Liquid-phase extraction, silica gel G column chromatography and thin -layer chromatography (TLC) were employed for sample fractionation. Th ree analytical high-performance liquid chromatography (HPLC) methods f or purification of NAEs were developed. Finally, analyses of NAEs by g as chromatography/mass spectrometry (GC/MS) are described. The applica tions of these analytical methods to the identification of anandamide and related NAEs in cell cultures as well as of artifacts in biosynthe tic studies are described.