Analysis of anandamide, an endogenous cannabinoid substance, and of other natural N-acylethanolamines
Abstract
Recent reports have suggested that N-arachidonoylethanolamine (anandamide) acts as an endogenous ligand for cannabinoid receptors in mammalian brain. Here we describe methods for the extraction, purification and 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. Three analytical high-performance liquid chromatography (HPLC) methods for purification of NAEs were developed. Finally, analyses of NAEs by gas chromatography/mass spectrometry (GC/MS) are described. The applications of these analytical methods to the identification of anandamide and related NAEs in cell cultures as well as of artifacts in biosynthetic studies are described.
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