Fontana A, Di Marzo V, Cadas H, Piomelli D
Istituto per la Chimica di Molecole di Interesse Biologico, C. N. R., Naples, Italy.
Prostaglandins Leukot Essent Fatty Acids. 1995 Oct;53(4):301-8. doi: 10.1016/0952-3278(95)90130-2.
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.
最近的报告表明,N-花生四烯酰乙醇胺(花生四烯酸乙醇胺)作为哺乳动物大脑中大麻素受体的内源性配体。在此,我们描述了花生四烯酸乙醇胺及相关N-脂肪酰基乙醇胺(NAEs)的提取、纯化和分析方法。采用液相萃取、硅胶G柱色谱和薄层色谱(TLC)进行样品分级分离。开发了三种用于纯化NAEs的分析型高效液相色谱(HPLC)方法。最后,描述了通过气相色谱/质谱(GC/MS)对NAEs进行分析的方法。阐述了这些分析方法在细胞培养物中花生四烯酸乙醇胺及相关NAEs鉴定以及生物合成研究中假象鉴定方面的应用。